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US 20070259939Al

(19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0259939 A1 Stebbing (43) Pub. Date: NOV. 8, 2007
(54)
USING NALTREXONE AS A MULTI-PURPOSE HEALTH SUPPLEMENT TO IMPROVE THE HUMAN CONDITION AND PREVENTING MULTIPLE DISEASES AND INFIRMITIES BY STIMULATING IMMUNE SYSTEM VITALITY AND ROBUSTNESS Publication Classi?cation

(51)

Int. Cl. A61K 31/485

(2006.01)

(52)

us. c1. ..................................................... .. 514/410

(75) Inventor:

Franklin Leroy Stebbing,

(57)

ABSTRACT

Norfolk, NE (U S)
Correspondence Address:
BANNER & WITCOFF, LTD. TEN SOUTH WACKER DRIVE, SUITE 3000 CHICAGO, IL 60606

Naltrexone has been used to ?ght existing infections, ai?ic

tions, and substance abuse. Embodiments of the current invention provide naltrexone as a preventative medicine

taken daily, usually in the late evening for greater efficacy.

It is taken in a loW dose like a vitamin supplement and may

prevent many diseases by stimulating the immune system.

The increased endorphin levels resulting from the use of loW dose naltrexone may also reduce the need for pain relief

(73) Assignee:

ACCELERATED TECHNOLOGIES, Norfolk, NE

medications, plus counter tendencies toWard experiencing

undesirable moods, such as depression or anger. Increased

(Us) (21) Appl. No.: (22)

Filed:

11/550,742
Oct. 18, 2006

feelings of Well-being produced by naltrexone may also have far-reaching eifects, as suggested by studies shoWing
that people With positive attitudes recover more quickly from serious illnesses, for example. In another aspect of the invention, naltrexone may be used to prevent substance addiction before it ever starts, by blocking the internal positive response that normally results from the use of chemical substances.

Related US. Application Data

(60) Provisional application No. 60/797,587, ?led on May

4, 2006.

US 2007/0259939 A1

Nov. 8, 2007

USING NALTREXONE AS A MULTI-PURPOSE HEALTH SUPPLEMENT TO IMPROVE THE HUMAN CONDITION AND PREVENTING MULTIPLE DISEASES AND INFIRMITIES BY STIMULATING IMMUNE SYSTEM VITALITY AND ROBUSTNESS CROSS REFERENCE TO RELATED APPLICATIONS

blocking the receptors that provide the stimulatory effect felt

from alcohol. As a result of the lack of positive reinforce ment, there is no internal incentive to drink alcohol. Naltr exone is not knoWn to have any important side effects at loW

[0001] This application claims priority to US. Provisional Application No. 60/797,587 ?led May 4, 2006.
FIELD OF THE INVENTION

dose levels. The FDA approved dosage siZe for treatment of alcohol dependency is a 50 mg tablet, once daily. While taking naltrexone at this dosage level, it is suggested that a counselor should be involved for support and advice and supervision. It also can be given in a sloW release form by injection or surgical implant if the patient is incapable or cannot be depended upon to take the medicine regularly.
SUMMARY OF THE INVENTION

[0006]

Previously, naltrexone had only been used to ?ght

[0002]

The invention relates to health supplements, in

existing infections or af?ictions, or to prevent recurrence. In

particular supplements for the prevention of diseases and

in?rmities and for the reduction of susceptibility toWards chemical addiction.
BACKGROUND OF THE RELEVANT ART

contrast, embodiments of this invention provide naltrexone as a preventative medicine taken daily, usually in the late evening for greater ef?cacy. It is taken like a vitamin supplement and may prevent many of the diseases that tend
to be susceptible to a strong immune system.

[0007]
[0003] Traditional medicine has long been focused on the cure of a?lictions after they are recognized by the onset of symptoms. Aspects of this invention are dedicated to the proposition that it is better to prevent diseases then to cure
them after they have become established. There are numer ous health supplements and vitamins available noW that are

The increased endorphin levels resulting from the

use of loW dose naltrexone may also reduce the need for pain
relief medications as Well as counter tendencies toWard

experiencing undesirable moods, such as excessive depres sion or anger. Increased feelings of Well-being may also

have far-reaching effects, as suggested by studies shoWing

that people With positive attitudes recover more quickly from serious illnesses, for example. In addition, any pre vention of sickness Would have the additional bene?ts of decreasing the use of medications With unWanted side effects
and loWering health care costs.

just for the maintenance and/ or enhancement of good health and vitality. More and more people are desirous of holding onto vigor and vitality as they age, such as baby boomers. Many people are determined to maintain their health and vitality for as long as it is possible to do so, and in?rmity is
often not accepted as inevitable. [0004] Naltrexone has been used in loW dose form since at least the mid 1980s to cure or stabiliZe a groWing list of ailments. It is used for the treatment of a number of serious diseases, such as some cancers, HIV, AIDS and multiple sclerosis. Studies have shoWn naltrexone to have a positive effect in as many as 65% of cancer patients, for example

[0008] In aspects of the invention, naltrexone may be used to prevent substance addiction before it ever starts, by taking it daily. This could result in great personal and ?nancial savings by reducing the occurrence of addiction and thus the need for rehabilitation programs. Unlike existing medica
tions such as Antabuse, a drug that Will induce nausea or

even unconsciousness if it is taken folloWed by alcohol consumption, naltrexone is more effective When taken prior
to drinking alcohol because it reduces or removes the desire to drink.

stopping the spread of tumors, as Well as causing remission in 30% of the patients. LoW dose naltrexone functions by blocking opioid, or endorphin, receptors in various types of

cells. By blocking opioid receptors, naltrexone also blocks

the reception of the opioid hormones that our pituitary and

adrenal glands produce: beta-endorphin and metenkephalin.

Many body tissues have receptors for these endorphins and enkephalins, including virtually every cell of the bodys
immune system. When the receptors are blocked, the body signals an increase in endorphin production and the level of endorphins rises in reaction. An increase in endorphins
causes the immune system to become stimulated and there fore more effective at ?ghting infection. The increased

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

[0009]

One embodiment of the invention is a method of

using loW doses of naltrexone to block endorphin receptors

in various body cells of a human or animal patient. When the receptors are blocked, the body signals an increase in

endorphin production and the level of endorphins rises in

reaction. The method may comprise a daily dose of naltr exone administered to the patient, for example betWeen

endorphin level may also dull or kill pain, and often results in a feeling of Well-being because endorphins are natural mood elevating chemicals in the brain. In contrast, stress
may cause a marked reduction in endorphins, Which often

approximately 0.5 and 5 milligrams of naltrexone, and preferably betWeen 1.75 and 4.5 milligrams of naltrexone. An aspect of the invention provides administration of the daily dose of naltrexone in the late evening hours, such as
betWeen 6:00 pm and 2:00 am, so as to block the opioid

does not correct itself naturally, thus resulting in loWered immune defenses susceptible to a variety of illnesses, such
as autoimmune diseases or cancer. Cancer has occasionally

receptors at a time When endorphins are naturally produced

by the body s circadian rhythm, usually in the early morning

betWeen 2:00 am and 4:00 am, regardless of Wakefulness or

been observed in patients after about tWo to six years folloWing a period of extreme stress. [0005] With respect to the treatment of alcohol addiction, naltrexone may be effective because it eliminates the neural

sleep. The brief blockade of opioid receptors betWeen 2 am.

and 4 am. may produce a prolonged stimulation of the

positive reinforcement normally experienced in the brain by

immune system by causing an increase in endorphin and enkephalin production. In fact, tests have shoWn that taking

US 2007/0259939 A1

Nov. 8, 2007

small doses of naltrexone each night has resulted in much

feeling of Well-being, naltrexone may reduce anti-social

behavior With more incidences of positive social interac tions. [0013] Naltrexone is already knoWn to help cure addictive

higher levels of beta-endorphins circulating in the blood in

the following days, providing as much as a 200-300%

increase in patients Who initially had de?cient endorphin

levels. The naltrexone may be taken as a tablet, capsule or

diluted in liquid, as long as it is present in a readily bioavailable form. [0010] An embodiment of the invention is a method to stimulate the immune system of a patient prior to the onset of an illness via a daily dose of naltrexone that is generally in the range of 0.5 to 5 milligrams per dose, and again

behaviors, such as alcoholism, opiate, narcotic and tobacco addiction. With respect to the treatment of alcohol addiction,
naltrexone may be effective because it eliminates the neural

positive reinforcement normally experienced in the brain by blocking the receptors that provide the stimulatory effect felt
from alcohol. As a result of the lack of positive reinforce
ment, there is no internal incentive to drink alcohol. In a

preferably betWeen about 1.75 and 4.5 milligrams per dose.

The resulting increase in endorphins thus causes a stimula

similar manner, in another embodiment of the invention, the

daily administration of loW dose naltrexone may greatly

reduce or eliminate the tendency to engage in addictive

tion of immune system function With marked improvement in disease ?ghting capability. In an aspect of the invention, the administration of a daily dose of naltrexone, for example
in the late evening, may prevent the onset of an illness in a

behavior by preventing its onset. This may be accomplished

by diminishing any potential internal reWards for using

chemical substances in the ?rst place. The loW dosage of naltrexone, for example around 4.5 mg, has been found to greatly inhibit alcohol cravings. The diminished desire for alcohol resulting from a loW dose of naltrexone may be effective for more than 24 hours, thus for example a dose
administered at 7:00 pm one evening may continue to inhibit

human or animal patient. The method includes taking loW

doses of naltrexone prior to potentially being exposed to

infectious conditions, With or Without additional medica tions or vaccinations to cause antibody production. An

increase in endorphins resulting from the blockage of endor phin receptors by the naltrexone causes the immune system
to become more effective at ?ghting infection during its early stages. The use of naltrexone as taught here may

alcohol cravings throughout the folloWing evening. Aperson

in need of curbing an alcoholic appetite may take a pill, either before or along With the drink, and discover no desire
for a second or third drink. To further eliminate an alcoholic

improve immune system function, resulting in a greatly

enhanced ability to fend off debilitating diseases such as

appetite, a series of 1.5 mg naltrexone doses may be taken

many forms of cancer, AIDS, HIV, multiple sclerosis, ALS, AlZheimers Disease, chronic fatigue syndrome, emphy sema, Parkinsons Disease, primary lateral sclerosis, psoria sis, rheumatoid arthritis, sarcoidosis and systemic lupus. For
example, cancer cells are routinely produced in the body, but a healthy immune system generally kills them as they occur. In fact, endorphins added to groWing human cancerous
tissue in a laboratory Petri dish have been observed to kill
the cancerous tissue. In contrast to Waiting for a disease to

Without alcohol, starting for example at about 6:00 pm. Three doses, for a total of 4.5 mg naltrexone, may be taken,
for example tWo or three doses together or single doses over a period of time, such as one per hour, as provides the most effective result for the individual. [0014] It is knoWn that it is very important to have a

positive attitude and outlook When battling and treating

serious illnesses, such as cancer. The outcome is much better

if the patient is upbeat and optimistic. This is similar to What

happens to those Who take loW doses of naltrexone to boost endorphin levels to combat diseases. The loW doses of

become established and then taking steps to cure it With

medications like naltrexone and/or other means such as

naltrexone raise the level of endorphins, Which eventually

leads to an increase in immune system function. Either Way,

surgery or chemotherapy, the disease may be prevented. Additionally, if an illness does become established, the daily naltrexone therapy may reduce or eliminate the need for
various medications to overcome the illness because of the

naturally or naltrexone induced, the increased levels of endorphins result in a better mood and an improved, enhanced and more robust immune system function, Which equates to an improved prognosis due to a greater ability of
the immune system to overcome and defeat a disease. If a

enhanced natural ability of the immune system.

[0011] An additional aspect of the invention is a method to alleviate or reduce pain in a human or animal patient comprising a daily administration of loW dose naltrexone. A naltrexone dose of betWeen about 0.5 and 5 milligrams may

patient does not have a naturally optimistic attitude, naltr

exone may help provide a good disposition With the same

bene?cial results, by arti?cially stimulating the immune system.

[0015] Variations and modi?cations of the foregoing are Within the scope of the present invention. It should be understood that the invention disclosed and de?ned herein
extends to all alternative combinations of tWo or more of the individual features mentioned or evident from the text. All

be used, for example taken during the late evening such as

from 6:00 pm to 2:00 am, as described above. The increased

endorphin level may dull or kill pain, as endorphin levels naturally increase When injury occurs. Because naltrexone stimulates an endorphin increase, naltrexone therapy may reduce aches and pains. [0012] The employment of a daily loW dose of naltrexone
may prevent one or more psychological problems in a

of these different combinations constitute various alternative aspects of the present invention. The embodiments

described herein explain the best modes knoWn for practic

ing the invention and Will enable others skilled in the art to
utiliZe the invention. The claims are to be construed to

human, in an embodiment of the invention. This is due to the

resulting increase of endorphins, Which are natural mood elevating chemicals in the brain. An increased level of endorphins may preclude or prevent the onset of some mental depression or anger, for example, because endor phins are natural anti-depressives and mood enhancers. As a result of the absence of depression and improved mood and

include alternative embodiments to the extent permitted by the prior art. For example, endorphins may be administered
directly Without the use of naltrexone such as in those cases

Where a patient is not able to produce endorphins by

naltrexone stimulation, or Where the use of naltrexone is

US 2007/0259939 A1

Nov. 8, 2007

prevented by other factors. For instance, because naltrexone

is an opiate agonist, it may not be appropriate to administer

naltrexone to a patient using opiates for pain medication.

[0016] Various features of the invention are set forth in the

following claims.
What is claimed is:
1. A method to increase endorphin levels in a human or

animal patient comprising a daily administration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 2. The method according to claim 1 Wherein the naltrex one is administered to the patient during an evening hour. 3. The method according to claim 2 Wherein the evening hour is betWeen approximately 6:00 pm and approximately
2:00 am.

11. The method according to claim 9 Wherein the chemi cal substance is an opiate. 12. The method according to claim 9 Wherein the chemi cal substance is alcohol. 13. The method according to claim 9 Wherein the chemi cal substance is nicotine. 14. A method to reduce physical pain in a human or animal patient comprising a daily administration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 15. The method of claim 14 Wherein the naltrexone is

administered to the patient during an evening hour. 16. The method of claim 15 Wherein the evening hour is betWeen approximately 6:00 pm and approximately 2:00
am.

4. A method to stimulate the immune system of a human or animal patient prior to the onset of an illness comprising a daily administration of a dose of naltrexone. 5. The method according to claim 4 Wherein the dose of naltrexone is betWeen about 0.5 mg and about 5 mg. 6. The method according to claim 4 Wherein the naltrex one is administered to the patient during an evening hour. 7. The method according to claim 6 Wherein the evening

17. A method to prevent the onset of illness in a human or

animal patient comprising a daily administration during an

evening hour of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 18. The method of claim 17 Wherein the evening hour is

betWeen approximately 6:00 pm and approximately 2:00

am.

hour is betWeen approximately 6:00 pm and approximately

2:00 am.

19. A method to prevent one or more psychological

8. Amethod to prevent addiction of a human or an animal

problems in a human patient comprising a daily adminis

tration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone.
20. The method of claim 19 Wherein the one or more

patient to a chemical substance comprising a daily admin

istration of a dose of naltrexone.

9. The method according to claim 8 Wherein the naltrex

one is administered prior to any use of the chemical sub
stance.

psychological problems are selected from the group com

prising depression, anger and anti-social behavior.

10. The method according to claim 8 Wherein the dose of naltrexone is betWeen about 0.5 mg and about 5 mg.

US006288074B1

(12) United States Patent

Bihari

(10) Patent N0.:

(45) Date of Patent:

US 6,288,074 B1
Sep. 11, 2001

(54) METHOD OF TREATING

LYMPHOPROLIFERATIVE SYNDROME

Primary ExaminerDWayne C. Jones

Assistant ExaminerC. Delacroix-Muirheid

(76)
(*)

Inventor:
Notice:

Bernard Bihari, 29 W. 15th St., New

(74) Attorney Agent Or Firm_W?liam J Daniel

York, NY (US) 10011

Subject to any disclaimer, the term of this patent is extended or adjusted under 35

(57)

ABSTRACT

Lymphoproliferative syndrome, including such diseases as

U~S~C~154(b)by0dayS(21) AppL NO: 09/439,987

(22) F1 d 1 e :
(60)

malignant lymphoma, chronic lymphocytic leukemia, Hodgkins lymphoma, and non-Hodgkins lymphoma, is
treated in human patients by the administration by a phar
macologically effective mode or route of an essentially pure

NOV. 15 1999

Related U-S- Application Data

Provlslonal apphcanon NO 60/108328 ?led on NOV 17 1998. 7
"""""""""""""

opiate receptor antagonist, typi?ed by Naltrexone and

Naloxone, exerting substantially higher blocking action for Mu opiate receptor sites than against Delta opiate receptor
sites at a loW dose concentration Which produces therapeutic

( )
(58)

- '

' """""""""""" " 514/286; 514/281; 514005;

514/280, 281, 282, 908

results corresponding to those obtained by the administra

non of Naltrexone at a 10W dosage level 1n the range of 1'0

Field of Search ................................... .. 514/277, 279,

mg. to 10 mg. and at Which Delta receptor blocking activity

is at most small and Mu receptor blocking activity is

56 ( )

_ R f Ct d e erences l e U'S' PATENT DOCUMENTS

5,631,263 * 5/1997 Portoghese et al. ............... .. 514/279

signi?cant and most preferably substantially exclusive. Nal trexone is suitable for oral administration and is preferred.

* cited by examiner

6 Claims, N0 Drawings

US 6,288,074 B1
1
METHOD OF TREATING LYMPHOPROLIFERATIVE SYNDROME
CROSS-REFERENCE TO PROVISIONAL APPLICATION

2
patents despite the totally different nature of the earlier
diseases Which did not involve any malignant behavior. It is not possible to provide any coherent rationale for the therapeutic effectiveness of an essentially pure opiate recep

tor antagonist against the diseases falling Within the lym

This application is a complete application of my provi sional application Ser. No. 60/108,828, ?led Nov. 17, 1998.
SPECIFICATION
10

pholiferative syndrome. Indeed, any therapeutic activity of these antagonists for the lymphoproliferative syndrome
Would appear from general principles of immune action to be highly unlikely if not entirely out of the question. It is generally believed that the immunocompetent action of lymphocytes arises out of some surface structural feature, usually called receptor, that is capable in some Way of

This invention relates to the treatment of lymphoprolif

erative syndrome and is concerned speci?cally With treat ment of this syndrome, including by such diseases as malig

nant lymphoma, chronic lymphocytic leukemia, Hodgkins lymphoma, and non-Hodgkins lymphoma, by administra
tion of an essentially pure opiate receptor antagonist such as
Naltrexone and Naloxone at a loW level dosage.
BACKGROUND

15

recognizing, perhaps by capture, some structural feature of an invading foreign antigen. This recognition mechanism, Whatever its nature, initiates the production of antibodies effective against the particular antigen. Even assuming that lymphocytes have receptors for opiates, it is quite puZZling that blocking of these receptors by an opiate
antagonist Would in some Way interfere With Whatever is the causative factor or factors in lymphatic cancers. If an occurrence of the lymphoproliferative syndrome could be traced strongly to viral or bacterial action and if such action is assumed to involve receptor sites on

The use of an essentially pure opiate receptor antagonist in the treatment of several diseases has already been dis
closed in patents in Which I am named as an inventor. In US.

20

Pat. No. 4,888,346, issued Dec. 19, 1989, the treatment Was

for the acquired immune de?ciency syndrome (or AIDS) in

any of its knoWn states, including AIDS-related complex. In US. Pat. No. 5,013,739, issued May 7, 1991, the disease treated Was chronic fatigue syndrome While in US. Pat. No. 5,346,900, issued Oct. 18, 1994, the disease Was chronic herpes virus infections. In the latter patent, examples of
treatment of multiple sclerosis Was also disclosed. For the treatment of all these diseases, the amount of the

lymphocytes, one could possibly theoriZe that an opiate

25

receptor antagonist might be preferentially blocking such

sites and preventing any connection With viral or bacterial particles. HoWever, viral and bacterial sources are usually accepted as having a limited association With any disease of

30

the lymphoproliferative syndrome and this theory thus could hardly apply to all of the diseases of this syndrome.
In any case, While such an association might explain hoW administration of an opiate receptor antagonist could reduce the likelihood of the original initiation of the malignancy, it Would not explain hoW such administration could provide a therapeutic action after the malignancy has progressed to the
point of treatment. It is true for lymphatic cancers as for all cancers that at the time diagnosis is possible, the cancer is

essentially pure opiate receptor antagonist Was required to

be at a quite loW level corresponding in results to those obtained by the administration of Naltrexone at a dosage level of from 1.0 mg. to 10 mg., preferably at a dosage level of 1.0 mg. to about 5 mg., and most preferably up to about 3.0 mg. At dosage levels above about 10 mg., not only Were the desired therapeutic results not obtained but the effect of
35

necessarily at a Well-developed, if not advanced, stage of

40

the treatment appeared to be negative in acerbating the

disease.
I have noW discovered that administration of an essen

development. Receptor site preclusion cannot therefore

explain an interference With a malignant proliferation of the lymphocytes. This is particularly true since the presence of the opiate receptor antagonist does not appear to block

tially pure opiate receptor antagonist gives desirable and

bene?cial therapeutic results in the treatment of a group of

normal proliferation of the lymphocytes in the system and

45

closely related malignancies knoWn as lymphoproliferative

their differentiation into B- and T-type cells as is essential to

a viable immune function.

syndrome, Which includes malignant lymphoma, chronic

lymphocytic leukemia, Hodgkins disease (or Hodgkins

lymphoma), and non-Hodgkins lymphoma. As the generic
name suggests, these diseases are characteriZed by the

McLaughlin et al have disclosed in US. Pat. No. 4,689,

multiplication or proliferation of tissue of the lymphatic

332; among others, that opiate receptor antagonists. exem pli?ed by Naltrexone and Naloxone, When administered to a variety of living organisms, including tissue and cells, can
exert either a groWth accelerating or a groWth inhibiting

system, especially lymphocytes (cells) produced in the

lymph nodes. Lymphocytes are important components of the
human immune response system and upon exposure to a

foreign antigen in the human body naturally proliferate or

multiply to combat the antigen. In this group of
55

effect dependent upon the length of time the opiate receptor sites of the organism are completely and continuously blocked or occupied by the antagonist. Speci?cally, a groWth
accelerating or promoting action occurs With blockage for a period of at least 12 hours per day, as can be achieved With a dosage of at least about 10 mg. and preferably about 20 mg. per day, Whereas a groWth inhibiting action occurs With

malignancies, the proliferation goes out of control, resulting

in an abnormal level of lymphocytes in the blood stream, enlargement of the lymph nodes due to accumulation of the

lymphocytes there, and other symptomatic characteristics.

In the past, these diseases Were generally treated by radiology and chemotherapy often employing a combination of antineoplastic agents. I have discovered that the lym phoproliferative syndrome can respond to an entirely dif
60

blockage for only a period of about 2 to 12 hours per day,

as can be achieved With a dosage of less than 10 mg. doWn

to about 0.1 mg. per day. The groWth accelerating embodiment is said to be effec tive to proliferation, migration, and differentiation of cer
65

ferent therapeutic approach involving the administration of

essentially pure opiate receptor antagonists at the same relatively loW quantitative level that Was found useful for the treatment of the disorders disclosed in the above-speci?ed

tain speci?c cells or tissue, including organ tissues, neural tissue, bone marroW, red blood cells, lymphocytes, liver cells, etc. (See patent, col. 9, lines 1525 This disclosure,
hoWever, is silent as to Whether these any of these same

US 6,288,074 B1
3
cells or tissues can be subject to the growth inhibiting

4
nism by Which the body attacks and controls cancers gen erally. It is the lymphocytes, or their derivatives, Which recogniZe the foreign antigenic nature of cancer cells or of
antibodies associated thereWith and attack the cancer cells.

embodiment, the focus of the inhibiting embodiment dis closure paralleling the above excerpt being on Weight loss of the organism as a Whole. (See col. 9, lines 3245 and col. 10,

lines 4559.)
In addition, the groWth inhibiting aspect is described as
related to the prevention, treatment and control of cancer

The route set forth by McLaughlin et al. for controlling cancer by their treatment, as quoted above, does not make
sense When it is the lymphocytes themselves that are malig

(see col. 10, line 60col. 11, line 47), the essential disclo
sure in this connection being as folloWs; The action of the compounds of the invention can be employed to terminate
10

nant. HoW can proliferation of malignant lymphocytes be

inhibited Without at the same time inhibiting the natural

the rapid groWth patterns of cancer and related abnormali ties. It should be understood, that a regime of the present
compounds cannot dissipate or reduce a tumor mass or other

proliferation of normal lymphocytes essential to effective functioning of the immune system? Further, if the object of
the treatment is, as stated, to reduce the extent of the cancer

metastasiZed groWth. These compounds can only terminate the groWth of the abnormal cells and inhibit the continued

to a degree Within the capacity of the immune system (lymphocytes) to control, hoW does an active normal

lymphocyte recogniZe a malignant lymphocyte).

15

groWth thereof. HoWever, by preventing tumor groWth to

continue [sic], i. e. reducing the tumor burden, the bodys
oWn defense mechanism, i. e. the immune system, has the

For at least these reasons, therefore, the discovery that

essentially pure opiate receptor antagonists, such as naltr

exone or naloxone, exert therapeutic action against the

opportunity to rid the body of the cancerous groWth Whether benign or malignant. This aspect of the invention is particu

lymphoproliferative syndrome is believed to be unexpected

if not remarkable.
20

larly signi?cant in light of early diagnostic techniques Which

do reduce tumor siZe, or With procedures for tumor excision. Moreover, naloxone, naltrexone, or the other related com
pounds can be administered as a prophylaxis to human or

The diseases usually identi?ed Within the lymphoprolif erative syndrome have been previously identi?ed and their
symptoms and other characteristics can be found in abbre viated form in any medical encyclopedia, such as Miller

animal subjects Who may be exposed to potentially carcin

Keane Encyclopedia & Dictionary of Medicine, Nursing, &

25

genic [sic] agents.

The only cancer speci?cally identi?ed for treatment in McLaughlin et al. patent is neuroblastona in mice, as illus

Allied Health, 5th Edition, 1992, W. B. Saunders Co. or in

more expanded form in any medical text, such as for

example Cecil Essentials of Medicine, by Andreoli etal,

Copyright 1986 by W. B. Saunders Co. At present, the four
diseases identi?ed are considered to constitute the lymphop

trated by speci?c examples 2, 3, and 5, the only examples

special type of cancer, occurring in the sympathetic autono
mous nervous system, i. e. mainly the nerves of the spinal

concerned With cancer. Neuroblastoma is a rather rare and 30 roliferative syndrome but if other diseases should be char

acteriZed by excessive proliferation of lymphocytes, they

column, and is essentially limited to young children up to about 10 years of age. Evidence has been reported that neuroblastoma is peculiarly susceptible to natural immuno

too Would respond to the present therapy and should be deemed Within the scope of the invention. Similarly, While Naltrexone and Waloxone are presently
35

the only essentially pure opiate receptor drugs knoWn to

have received government approval for administration to humans, if other drugs exist or should be developed exhib
iting the same preferential or selective affinity for Mu over

biological resistance. According to IMMUNOBIOLOGY FOR THE CLINICIAN by Barber, Copyright 1977, John Wiley & Sons, the lymphocytes of disease-free mothers of young neuroblastoma patients have the capacity to kill
neuroblastoma cells extracted from their diseased children
but had no effect on normal tissue cells or tumor cells of
40

Delta opiate receptor sites, they too should be effective for

purposes of the present method and are Within the scope of the invention. For more details of this selective or prefer ential action, reference may be had to US. Pat. No. 5,013,

other than neuroblastoma and that lymphocytes of siblings

of such patients also Were able to kill neuroblastoma cells of

the patients. (Cf p. 69). The same text states (at p. 102) that incidence of neuroblastoma nodules found by autopsy is
4050 times greater than the overall incidence of clini

45

739, the relevant portions of Which, in particular col. 5, line 17col. 6, line 25, are incorporated by reference. Similarly, While the dosage levels have been brie?y
speci?ed above, more complete information as to dosage level Which is applicable to the present invention, is given in the same -739 US. patent, especially col. 6, line 59col. 7,
line 17, Which is incorporated by reference. In as much as Naltrexone is available in a form suitable for oral adminis tration and is recogniZed to be effective When so

cally diagnosed neuroblastoma. This obviously suggests that

a competent immune system exerts a strong controlling action on neuroblastoma Which is found in feW other kinds
of cancers.

While there is a tendency among the general public (and even the often equally uninformed media) to generaliZe

betWeen all cancers, (a conception Which unfortunately

appears to be shared by McLaughlin et al) cancer is, in
fact, a collection of many distinct malignancies, each With
55

administered, it is preferred that the Naltrexone be utiliZed as the opiate antagonist and be administered orally, but
Where effective other administration routes are, in principle, not precluded and can be employed. Naloxone, on the other hand, has not generally proven to be effective When admin istered orally; it is available in a form suitable for injection

its unique characteristics, behavior, and treatment response

(Which is one reason for the tortuously sloW progress in the treatment of cancer), and this individualistic nature is especially applicable in the case of neuroblastoma. One

cannot, therefore, reasonably extrapolate from the response

of neuroblastoma to a given prophylaxis to a supposition that an entirely different type of cancer Will exhibit the same response and this conclusion is particularly valid in the

60

and is better administered by injection, It is also preferred that administration take place in the evening hours, and particularly at bedtime, since the action
of the antagonist appears to develop more strongly When the patient is sleeping and at rest.
EXAMPLES

case of lymphoproliferative malignancies.

As already noted above, lymphocytes are at least a core 65

Example 1
A37 year old female patient had been diagnosed 31/2 years earlier as having non-Hodgkins lymphoma and had been

component, if not the keystone, of the bodys immune system Which is an important, if not the principal, mecha

US 6,288,074 B1
5
successfully treated into remission With chemotherapy by an
oncologist in a different state. With this treatment, the

6
levels and his Weight loss regained, At this point, he Was
reevaluated by his oncologist and Was told he Was in remission. His Naltrexone therapy has continued for a total of about 3 years and he has remained free of all symptoms.

symptoms experienced during the active phase of her

disease, namely, tumor masses and associated Weight loss,

fevers, loW energy, life-threatening systemic infections, and

malaise, had all cleared and she enjoyed this remission for

During this time, X-rays at 6 month intervals and CT Scans

at 12 month intervals have remained normal.

three years. Then, hoWever, lymphoma reappeared in lymph

glands in her neck, chest, and abdomen and her earlier

Example 3
10

symptoms of fevers, malaise, Weight loss, reduced energy,

and increased vulnerability to systemic infections recurred. Lymph node biopsy Was positive for lymphoma and che
motherapy Was reinstituted. She did not respond to chemo therapy and her disease Was judged to be resistant to this

AWoman Was examined and found to have three golf-ball siZe nodes in her groin Which on biopsy Were positive for

non-Hodgkins lymphoma. When she refused

chemotherapy, she Was started on Naltrexone at a dosage of

3 mg. by mouth qhs (i. e. every night at bedtime) and after

15

therapy. Six months after this recurrence began, her Weight loss had reached 20 pounds, she had enlarged lymph nodes in her axilla and groin, an enlarged spleen, an elevated White blood cells count With a number of atypical lymphocytes, and Was naturally quite frightened at her condition.
She Was started on Naltrexone at 3 mg. by mouth every

12 Weeks her groin nodes disappeared. She continued on this Naltrexone treatment and remained free of nodes until her death four years later from arteriosclerotic heart disease

(ASHD).
What is claimed is:

1. A method of treating a human patient suffering from

night at bedtime. After 3 Weeks, her fever began to subside and after 3 months, all of her symptoms had cleared Without
treatment other than the Naltrexone. After about 4 months of

lymphoproliferative syndrome, Which comprises the step of

administering by a pharmacologically effective mode to said patient a therapeutically effective dose of a therapeutic agent
25

Naltrexone treatment, she Was examined by her original oncologist and her related lab Work repeated, Which shoWed that her X-ray and blood abnormalities had cleared. She had regained her lost Weight, Was free of all other symptoms and
Was adjudged to be in full remission. The loW dose Naltr
exone treatment has been continued and after about one year,

consisting essentially of an essentially pure opiate receptor antagonist having a selectively higher blocking action against Mu opiate receptors than against Delta receptors, the
amount of said dose being selected to produce therapeutic

results substantially corresponding to those produced by

Naltrexone When administered in the range of about 1 mg to

she remains in remission.

Example 2
A 38 year old man had been diagnosed by an oncologist as having Hodgkins disease based on symptoms of enlarged cancerous lymph nodes in the paravertebral areas of his chest and abdomen, an enlarged spleen, poor appetite, a

about 10 mg per day. 2. The method of claim 1 Wherein said lymphoprolifera tive syndrome is selected from the group consisting of

lymphoma, chronic lymphocytic leukemia, Hodgkins lym phoma and non-Hodgkins lymphoma.
35

20-pound Weight loss, malaise and fever. After receiving radiation and chemotherapy, these symptoms cleared, his
Weight loss Was re-gained and he Was declared to be in remission. After about four years, his symptoms recurred

3. The method of claim 1 Wherein said antagonist is administered at bedtime of the patient. 4. A method of treating a human patient suffering from

lymphoproliferative syndrome, Which comprises the step of

administering by a pharmacologically effective mode to said patient a therapeutic agent consisting essentially of an essentially pure opiate receptor antagonist having a selec

including the enlarged spleen, enlarged paravertebral nodes

in his chest and abdomen as shoWn by X-ray, tiredness, loss of appetite, and Weight loss. He Was again treated With
chemotherapy With no response and he Was advised that his

tively higher blocking action against Mu opiate receptors

than against Delta receptors in an amount Which is effective

disease become resistant to chemotherapy.

Some 6 months subsequent to his reoccurrence and failed

45

to exert a substantially higher opiate blocking action against Mu receptors than against Delta receptors. 5. The method of claim 4, Wherein said opiate receptor
antagonist is either naltrexone or naloxone. 6. The method of claim 4 Wherein said lymphoprolifera

chemotherapy and folloWing a neW examination con?rming the diagnosis of Hodgkins lymphoma, he Was started on Naltrexone at 3 mg. by mouth every night at bedtime. After four months of taking Naltrexone at this dosage, his enlarged nodes and spleen shrank to normal siZe as revealed by X-ray ?lm and Ct scans, his energy and appetite returned to normal

tive syndrome is selected from the group consisting of

lymphoma, chronic lymphocytic leukemia, Hodgkins lym phoma and non-Hodgkins lymphoma.
* * * * *

US008067430B1

(12) United States Patent

Ugen et a].
(54) ANTI-HIV ACTIVITY OF THE OPIOID
ANTAGONIST NALOXONE

(10) Patent N0.: (45) Date of Patent:

US 8,067,430 B1
Nov. 29, 2011

Schmidt, William K. Alvimopan (ADL 8-2698) is a Novel Periph eral Opioid Antagonist. The American Journal of Surgery. 2001. vol. 182, Suppl to Nov. 2001). pp. 27S-38S.*
Suzuki et al, Methadone induces CCR5 and promotes AIDS virus infection, FEBS Letters, 519 , 2002, pp. 173-177.* Medicinenet (De?nition of HTLV-III, retrieved from the internet on

(75) Inventors: Kenneth E. Ugen, Tampa, FL (US); Steven Specter, Tampa, FL (US); Susan B. Nyland, Hershey, PA (US); Chuanhai

May 14, 2010, HTML: http://WWW.medterms.com/script/main/art.

Cao, Tampa, FL (US)

(73) Assignee: University of South Florida, Tampa, FL

asp?articleke}P38558.*
Schafer et al. Drug Discovery Today 2008, 13 (21/22), 9l3-9l6.* Horig et al. Journal of Translational Medicine 2004, 2(44).* Ando et al.(Remington: The science and practice of pharmacy, 20th

(Us)
(*) Notice: Subject to any disclaimer, the term of this patent is extended or adjusted under 35

Edition, pp. 704-712).*

Gekker, Ganya et al., Naltrexone Potentiates Anti-HIV-l Activity of Antiretroviral Drugs in CD4+ Lymphocyte Cultures, Drug and Alco hol Dependence, 2001, 257-263, vol. 64.

U.S.C. 154(b) by 1363 days.

Ho, Wen-Zhe et al., Methylnaltrexone AntagoniZes Opioid-Mediated

Enhancement of HIV Infection, The Journal of Pharmacology and

(21) App1.N0.: 10/902,471

(22) Filed: Jul. 29, 2004
Related US. Application Data

Exp. Therapeutics, 2003, 1158-1162, 307(3).

Gekker, G. et al. Jun. 2002. Naloxone PotentiatesAnti-HIV-l Activ

ity of Antiretroviral Drugs in CD4+ Lymphocyte Cultures. Interna

tional Journal of Infectious Diseases. vol. 6. Suppl. 2. pp. Sl4-Sl5. Gekker, G. et al. Jan. 2001. Naltrexone Potentiates Anti-HIV-l

(60)

Provisional application No. 60/481,156, ?led on Jul. 29, 2003.

Activity of Antiretroviral Drugs in CD4+ Lymphocyte Cultures. Drug and Alcohol Dependence. vol. 64. pp. 257-263.
Brown et al. 2009. Low-Dose Naltrexone for Disease Prevention

and Quality of Life. Medical Hypotheses. vol. 72. No. 3. pp. 333
337.

(51)
(52)

Int. Cl. A61K 31/485

Wang et al. 2007. Diffrerent Effects of Opioid Antagonists on Mu-,

(2006.01)

Delta-, and Kappa-Opioid Receptors With and Without Agonist

Pretreatment. The Journal of Pharmacology and Experimental
Therapeutics. vol. 321. No. 2. pp. 544-552.

US. Cl. ..................................................... .. 514/282

(58)

Field of Classi?cation Search ................. .. 514/282

See application ?le for complete search history.

(56) References Cited
U.S. PATENT DOCUMENTS
4,546,103 A * 10/1985 Huebner ..................... .. 514/282

(Continued)
Primary Examiner * Jeffrey S Lundgren
Assistant Examiner * Savitha Rao

(74) Attorney, Agent, or Firm * Michele L. LaWson; Smith

. 514/282

4,888,346 A *
2003/0069263 A1 *
2003/0105121 A1 *

12/1989 Bihari et al.

4/2003
6/2003

& Hopen, PA.

Breder et al. ............... .. 514/282

Bihari ......................... .. 514/282

(57)

ABSTRACT

OTHER PUBLICATIONS

A method of treating HIV infection by using the novel anti HIV activity of the opioid antagonist naloxone.
2 Claims, 4 Drawing Sheets
Fenzm lnm ' no! RTmMty

De Clercq, Eric. New Developments in anti-HIV chemotherapy.

Biochimica et Biophysica Acta. 2002. vol. 1587. pp. 258-275.*

A Function 01 N

mn- 'hlnmcm: now

aa

auRombTiMmtfoKn 68 3
5 g 5 n
w w in w u: w 10 H1 0

Molar 04) cunmnmlnm ul' Ndnmneml

Ema or AZT Ti-tmem M
FIV-I mum cu. an m- Auivmy

or n mMny Trutmenl mm

AMeusrnlmou'mn

nasi

RT - memo mmt?mue (Inlvily mnsllEd B mlllmltl/ml)

US 8,067,430 B1
Page 2
OTHER PUBLICATIONS
Peng et al. 2007. Pharmacological Propeities of Bivalent Ligands

NaloXone at Mu, Delta, and Kappa Opioid Receptors. J. Med. Chem.

V0
1.50.

pp

. 2254-2258.

Containing Butorphan Linked to Nalbuphine, NaltreXone, and

* Cited by examiner

US. Patent

Nov. 29, 2011

Sheet 1 of4

US 8,067,430 B1

F l GU R E I A- D
Effect of Naioxone Treatment of HIV-1 Infected Cells on RT Activity

Percent In hibition of RT Activity

as A Function of Naloxcna Treatment Doses

womtv
9387.1
10

"3

10

1e

16

1 o 10

ID

Molar (M) Concentrations of Naloxon e-HCI

Effect Of AZT Treatment of HIV-1 Infected Cells on RT Activity

200 190 180 170
160

Hc:.E*0ouEnmcs5gQ-2.E0#.EF5E32S4W967M54321 A

150 I40 130

120 I 10 I00 90

0amHw9B765M2l

.@ lun
t

sc

m an I

m a

Am N 40 w .m we m .Mm my Or aw PA . _ mm .m ..._.m 8 M .w \IH M . an m w er.IIU mm aw m

n D w

O h 0 s 0 O

Molar (M) Concentrauons of AZT

Moiar (M) Concentrations of AZT

RT - reverse transcriptase (activity measured as milIunIts/ml)

AZT =- azidothymldine (zidovudine)

mU/rnl - milliunitslml

SD - standard deviation

US. Patent

Nov. 29, 2011

Sheet 2 of4

US 8,067,430 B1

FIGURE 2A and B
Percent inhibition of Recombinant RT
' Activity as A Function of Naloxone

Effect of Naloxone Treatment

on Recombinant RT Activity
A

Treatment DOSES

MRT (A: SD ecmtuai/vnmit}y

10'11 161010 10 16

PlIof iRT Anehctbainvo!ty

16 166 164
1o
11

101

Q 0

1|]9 10

10

10

10

1o

Molar Concentrations of Naioxone-HCL

Molar coneantrations oi Naloxone-HCL

mU/mi = miliiunits/ml

individual value labels are shown above each bar

US. Patent

Nov. 29, 2011

Sheet 3 of4

US 8,067,430 B1

In

Effect of naloxone or AZT treatment of HIV-1 infected on

gp120 expression

Key:
Std = protein molecular weight standard of 98KB in size

US. Patent

Nov. 29, 2011

Sheet 4 of4

US 8,067,430 B1

FlGURE 4

MoiuOonomrations(M)>io w'5 s04 10 1o~8 i0" i6

of Naloxore

10

VC 0C "115'

RT-PCR amp?fication of HN-lgp4l (prominent

white bands in each lane) from HIV-MN infected Sup'l'i cells treated with naloxone at a range of molar concentrations.
VC == virus control (no treatment with drug) CC = cell control (cells not infected with HIV-1) Marker = DNA molecular weight markers of different sizes

US 8,067,430 B1
1
ANTI-HIV ACTIVITY OF THE OPIOID ANTAGONIST NALOXONE CROSS REFERENCE TO RELATED APPLICATIONS

2
In yet another embodiment, provided is an antiviral com

pound comprising an opioid antagonist, Wherein the opioid antagonist is naloxone hydrochloride.
BRIEF DESCRIPTION OF THE DRAWINGS

This application claims priority to US. Provisional Patent Application No. 60/481,156 of the same title and by the same inventors, ?led Jul. 29, 2003.
GOVERNMENT SUPPORT

For a fuller understanding of the nature and objects of the invention, reference should be made to the folloWing detailed description, taken in connection With the accompanying

draWings, in Which:
FIG. 1 (A-D) shoWs the results of the differential effect of
a range of molar concentrations of naloxone or AZT on

National Research Service Award (NRDA)4Grant No.

DA05913. BACKGROUND OF THE INVENTION

reverse transcriptase activity in HIV infected cells: FIG. 1A shoWs the RT activity in HIV-MN infected Supt 1 cells treated With different molar concentrations of naloxone.
The data demonstrate that as the treatment dose of naloxone

The pandemic ofAIDS (Acquired Immunode?ciency Syn

drome) represents one of the greatest challenges faced by biomedical science. Although AIDS has been kept in check,
to a certain degree, in the U. S. through the use of antiretroviral therapy, infections still occur and not all Americans have
20

increases from 10-11 to 10-4 M RT activity decreases. The column in the graph labeled 0 is the control (i.e. no drug

treatment).
FIG. 1B shoWs the % decrease in RT activity for the dif ferent treatment doses compared to the RT activity in the control.
FIGS. 1C and 1D shoW results from the treatment of HIV MN infected Supt 1 cells With an identical molar dose range
25

access to appropriate therapies. In other parts of the World,

particularly Africa, AIDS is devastating With infections

occurring at an unchecked rate. In addition, in those infected

of the established anti-HIV drug AZT.

FIGS. 2A and 2B summariZe the results of the effect of different molar concentrations of naloxone on recombinant

regions, sophisticated antiretroviral therapy is not available Which often results in high and often rapid mortality.
Short of the availability of an effective vaccine it is abso

RT (that is, protein not Within the HIV viral lysate). Analo
gous to the effects on RT in HIV, naloxone demonstrated a
30

lutely essential to develop neW, effective and economical drug

direct inhibitory effect on RT.

therapies. This is particularly important because of the ability of the human immunode?ciency virus (HIV), the causative
agent for AIDS, to become resistant to some effective thera

FIG. 3 shoWs the immunoblot (Western blot) results using lysates from HIV infected Supt 1 cells that had been treated
With different molar concentrations of either AZT or nalox
35

pies.
One of the problems With the generation of neW drug

therapies against HIV is the development of resistance by the virus to the therapy. Unfortunately, the development of resis
tance has become a serious problem With AZT (Zidovudine). Resistant viruses are also developing against the neWer pro

one. The expression of the essential HIV coat protein gp120 is measured by this technique. Western Blot analysis of gp 1 20 detected With a murine anti-gp 120 monoclonal antibody from
HIV-1 infected cells treated With naloxone or AZT. HIV-MN

tease inhibitor drug therapies. With the development of resis tance it is critical to continually develop neW drug therapies
against HIV. Therefore, What are needed are novel ef?cacious

40

infected Supt 1 cells treated Will different molar concentra tions of naloxone or AZT. Cells Were lysed at 3 days after infection of the cells With cell free HIV-1MN and 100 pg of
the lysate Was loaded onto SDS-PAGE, transferred onto a

Hybond ECL membrane and detected With an anti-gp120

drug therapies for the treatment of HIV infection and AIDS.

45

monoclonal antibody. VCq/iral control (infected cells With no drug treatment).

FIG. 4 is a RT-PCR analysis of RNA from HIV infected target cells Which had been treated With different molar con centrations of naloxone. The expression of the critical coat

SUMMARY OF INVENTION

This invention describes the discovery of novel anti-HIV activity for naloxone hydrochloride, NARCAN, Which is cur

protein gp41 is being measured by this molecular analysis.

50

rently used to treat opioid abuse (morphine, heroin, etc.).

In one embodiment the inventive method provides for the treatment of a patient, infected With a virus, comprising the

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

step of administering to the patient a therapeutically effective amount of an opioid antagonist. The target virus is the human

In the folloWing detailed description of the preferred

55

immunode?ciency virus (HIV) and the opioid antagonist is naloxone hydrochloride.

In another embodiment, provided is a method of inhibiting
reverse transcriptase activity in a cell infected With the human

immunode?ciency virus (HIV) comprising the step of con

tacting the infected With a therapeutically effective amount of
60

embodiments, reference is made to the accompanying draW ings, Which form a part hereof, and Within Which are shoWn by Way of illustration speci?c embodiments by Which the invention may be practiced. It is to be understood that other embodiments may be utiliZed and structural changes may be made Without departing from the scope of the invention. Administering or Contactingias used herein refers to the
process of delivering to a cell, ex vivo, or a host, in vivo, a

naloxone hydrochloride.
Another embodiment of the present invention includes a method of modulating the expression of a coat protein of a

virus comprising the step of contacting the virus With a thera peutically effective amount of naloxone hydrochloride,
Wherein the coat protein of the virus is chosen from the group

65

therapeutic sub stance, or a combination of several therapeutic substances. The process can include any method knoWn in the art and is dependent on the type of substance or substances administered. Possible methods include, but are not limited

to, parenteral (i.e. subcutaneously, intravenously, intramus

cularly, intra-arterially, and direct injection into a tissue or

consisting of gp41 and gp120.

US 8,067,430 B1
3
organ), mucosal (i.e. intranasally), pulmonary (i.e. via inha lation), topical, via catheter (i.e. iontopheretically) or orally.
Administration is usually achieved via a pharmaceutically

4
experiment, the effects ofAZT or naloxone (at identical molar concentrations) on RT activity. Naloxones Effect on Retrovirus Binding Activity
In addition to having a direct effect on RT activity the ability of naloxone to inhibit HIV Was measured through the determination of expression of critical coat proteins of HIV

acceptable carrier.
CCR-Sias used herein refers to the protein encoded by
the gene for CCR5 Which is located on chromosome 3 in the

p21.3-p24 region. CCR5 is a chemokine receptor present in

(called gp41 and gp120). This Was accomplished either

different cells, especially in macrophages, monocytes, and T

cells, Where it acts as a co-receptor for HIV-1 in these cells. Chemokines and their receptors are believed to be involved in

through RT-PCR (reverse transcriptase-polymerase chain

reaction) Which measures messenger RNA (mRNA) and by

immunoblotting (Western blotting) Which directly measures

the expression of protein. Naloxone treatment of HIV infected lymphocytes Was able to signi?cantly depress expression of these necessary proteins through measurement

the in?ammatory response, mediating leukocyte movement

and activation. CD4ias used herein refers to a T-cell receptor protein

Which interacts With and recogniZes major histocompatibility

complex proteins. CD4 is the receptor for the human immu node?ciency virus (HIV) and may mediate its entry into the host cell. It has a high a?inity for the envelope protein of HIV,

of mRNA and protein expression.

Example 1
Cell Infection by HIV and Sample Preparation for

gp160, and also for its split product, gp120.

gp120ias used herein refers to glycoprotein 120, a protein
that protrudes from the surface of HIV and binds to CD4+ T cells. In a tWo-step process that alloWs HIV to breach the membrane of T cells, gp120-CD4 complex refolds to reveal a second structure that binds to CCR5, one of several chemok
20

Analysis
200TCID-50 per milliliter (tissue culture infectious dose)
of HIV-MN cell free virus Was used to infect 500,000/milli

liter of a human T cell line (called SupTI) in the presence (at

25

ine co-receptors used by the virus to gain entry into T cells. gp41ias used herein refers to glycoprotein 41, a protein embedded in the outer envelope of HIV. gp 41 plays a key role in HIVs infection of CD4+ T cells by facilitating the fusion
of the viral and the cell membranes. Therapeutically Effective Amountias used herein is that
amount of a substance necessary to achieve a desired thera

a range of concentrations) or absence (control) of naloxone HC1/or AZT. The TCID-50 is an amount of virus Which Will infect 50% of non-infected target cells. The virus+cells+ drugs (naloxone or AZT) Were incubated at 37 degrees Cel sius in an atmosphere of 5% carbon dioxide for three days.

30

peutic result. For example, if the therapeutic result desired is the enhanced yield of stem cells, the therapeutically effective
amount is that amount that facilitates, or achieves, an increase

in the total number of stem cells in a given population. The therapeutically effective amount can be a dosage adminis
tered in at least one amount and can include an administration

35

Cells Were lysed by using a detergent 1% Triton-X100. This preparation Was froZen in liquid nitrogen and thaWed at 37 degrees Celsius for three cycles. The preparation Was vor texed vigorously for 20 seconds at the end of each cycle. Samples Were then stored at 80 degrees Celsius until analy sis. Total RNA from the samples Were prepared by using the Tri-reagent assay and aliquoted and stored at 80 degrees

Celsius until RT-PCR (reverse transcriptase-polymerase

chain reaction) analysis.

Example 2
40

protocol spanning several days or Weeks.

This inventive method includes the use of the novel anti

HIV activity of naloxone hydrochloride (NARCAN), a nar cotic antagonist, to treat patients infected With HIV. The medical indication for this drug is the treatment and partial, or total, reversal of the effects of opioid abuse. That is, naloxone is a competitive narcotic antagonist used in the management and reversal of overdoses caused by narcotics and synthetic narcotic agents. Unlike other narcotic antagonists, Which do

RT (Reverse Transcriptase) Activity Assay (Used to Mea

sure Effect of Drug Treatments on HIV Infected Cells) A 96 Well microplate Was coated With 200 ul/Well of a 40
45

mg/ml solution of polyA and incubated at room temperature overnight With rocking. The plate Was then Washed 3>< With

not completely inhibit the analgesic properties of opiates, naloxone antagoniZes all actions of morphine.
Naloxones Effect on Reverse Transcriptase Activity Many studies have dealt With the effects of opioids (mor phine and analogs) on infection of cells by pathogenic human retroviruses (HIV and another retrovirus called the human T cell leukemia virus (HTLV)). In these experiments it may be necessary to antagoniZe the effect of the opioids With an antagonist, in order to con?rm the speci?city of any effect by
50

TBS (Tris buffered saline, pH 7.8) 100 ul/Well of either lysed experimental samples (from infected drug treated cells) or RT
standards Were added to the Wells folloWed by the addition of 60 ul/Well of RT buffer. This Was folloWed by the addition of 40 ul/Well of a nucleotide mix. The reaction mixture Was then incubated at 37 degrees Celsius for 40 min. The plates Were then Washed 3>< With TBS folloWed by the addition of anti

DIG-POD-HRP (a speci?c antibody against RT Which alloWs

for the accurate measurement of this enzyme) at a dilution of
55

the opioids. Here, the opioid antagonist naloxone (NAR

CAN) Was chosen. Surprisingly, it Was discovered in control

115000 in TGSE (a 10 mM pH 7.5 Tris buffer containing 1% gelatin and 1 mM sodium EDTA) buffer at 100 OWen and
incubated for 1 hour at 37 degrees Celsius. Plates Were then

experiments using antagonist alone that naloxone treatment

exerted anti-HIV activity. This Was indicated by a direct

inhibitory effect by the drug on the reverse transciptase (RT) enZyme activity of the virus. RT activity is essential for rep lication of the virus and ultimately the ability of the virus to infect cells. By comparison, the established anti-HIV drug
AZT (Zidovudine) inhibits HIV through its effect on RT. Importantly; initial analysis indicates that naloxone is at least
as effective as AZT in inhibiting RT activity at similar con

60

Washed 6>< With TBS folloWed by the addition of 100 ul/Well of a soluble substrate for HRP designated TMB (tetra-methyl benZidine) in perborate buffer. Color development, due to the reaction of the substrate With the enZyme (HRP) conjugate
Was alloWed to occur at room temperature for 15 min. The

65

colorimetric reaction Was stopped by the addition of 25 ul/Well of 2 molar sulfuric acid. The color reaction in the Wells Was measured spectrophotometrically. Color development
Was proportional to the amount of RT in the Wells and quan titation Was made based upon a standard curve of using

centrations. This Was determined by measuring, in the same

US 8,067,430 B1
5
recombinant RT. FIG. 1 (A-D) shows the results of the dif
ferential effect of a range of molar concentrations of naloxone orAZT on reverse transcriptase activity in HIV infected cells.

6
protein gp4l is measured by this molecular analysis. As
shoWn in FIG. 4, naloxone inhibits HIV at the molecular level

by inhibiting expression of RNA for the envelope glycopro

tein gp4 1.

Speci?cally, FIG. 1A shoWs the RT activity in HIV-MN infected Supt 1 cells treated With different molar concentra
tions of naloxone. The data demonstrate that as the treatment

Example 4

dose of naloxone increases from 10'1 l to 10'4 M RT activity decreases. The column in the graph labeled 0 is the control (i.e. no drug treatment). FIG. 1B shoWs the % decrease in RT activity for the different treatment doses compared to the RT activity in the control. FIGS. 1C and 1D shoW results from the
treatment of HIV-MN infected Supt 1 cells With an identical

Immunoblot (Western Blot) Assay (for Quantitation

of Viral Protein Expression as a Function of

Treatment With the Drug Regimens)

100 mg of protein lysate (i.e. from the HIV infected cells)
from the samples described above Were loaded onto a SDS

molar dose range of the established anti-HIV drug AZT.

Accordingly, naloxone is at least as effective as AZT in inhib

PAGE (sodium dodecyl sulfate-polyacrylamide gel electro

phoresis) and then transferred to a hybond ECL membrane (considered the blot). This blot Was probed, folloWing an ECL kit protocol (Amersham Phamarcia, Inc.), With a mouse anti

iting RT activity in HIV infected cells.

FIGS. 2A and 2B summariZe the results of the effect of different molar concentrations of naloxone on recombinant

RT (that is, protein not Within the HIV viral lysate). Analo
gous to the effects on RT in HIV, naloxone demonstrated a direct inhibitory effect on RT. Therefore, it can be seen that

HIV-l gpl20 (external envelope glycoprotein). Binding of

the mouse monoclonal antibody to the gpl20 immobiliZed in the blot Was detected by an anti-mouse HRP (horseradish peroxidase)/ sub strate similar to the one described above. The absence of or a decreased intensity of the gpl20 band from infected cells treated With the drugs compared to control (non-treated) is indicative of an inhibitory effect of the drug
25 treatment on HIV.

naloxone inhibits HIV through its effect on RT.

Example 3
RT-PCR (Reverse Transcriptase-Polymerase Chain Reac
tion)i(for Quantitation of Viral RNA as a Function of Treat

ment With the Drug Regimens)

Total RNA from the preparations described above Was used as a template for the generation of complementary DNA (cDNA). This Was accomplished through the use of commer
30

FIG. 3 shoWs the immunoblot (Western blot) results using lysates from HIV infected Supt 1 cells that had been treated
With different molar concentrations of either AZT or nalox

one. The expression of the essential HIV coat protein gpl20

cial RT-PCR kits (Gibco/ BRL/ Life Technologies, Inc., Rock ville, Md.). The gene for the HIV gp4l transmembrane enve lope glycoprotein Was ampli?ed from the RT products through the use of the folloWing PCR cycling conditions: 94 degrees Celsius for 2 minutes folloWed by 52 degrees Celsius for 1 minutes then folloWed by 32 cycles at 94 degrees Celsius for 1 minutes, 52 degrees Celsius for 1 minutes, 72 degrees
Celsius for 15 minutes, then PCR extension Was accom

is measured by this technique. Thus, naloxone has been

shoWn to be as effective, or more effective, than equivalent molar concentrations of AZT in inhibiting HIV as measured

by expression of the HIV envelope glycoprotein.

35

There are several Ways to quantify the activity of antiviral

drugs. A direct method requires the ability to directly detect

and measure a suitable biological endpoint. Indirect measures

plished at 72 degrees Celsius for 10 minutes. The cycles Were then ended by incubation at 4 degrees Celsius for 16 hours. 5
ml (microliters) of PCR products Were then loaded onto a

40

1.5% agarose gel containing 5 ug/ml of ethidium bromide.

Bands in the gel With a siZe of 1.2 kb Was considered to be

include the effects of the drug on the expression of viral messenger RNA or protein necessary for infectivity at the gene and/ or protein. As for anti-HIV drugs, most recogniZed methods used are the measurement of HIV-I protein level

gp4l based upon the primers used in the assay: Primer

Sequences are as folloWs: Sense: 5 prime-CAGCGATAG

indicated by p24 (HIV-l core antigen) levels, RT (reverse transcriptase) activity, and immumoblot (i.e. Western)
method. The method for measurement of messenger RNA
45

GAGCTCTGTTCC-3 prime (SEQ ID NO:l); Antisense 5

prime-GACCATTTGCCACCCATCTTA-3 prime (SEQ ID

NO:2). In order to eliminate the possibility that the drug
treatments could have an inhibitory effect on the RT used in the RT-PCR reaction rather than the HIV RT a constitutive (i.e. alWays expressed) enZyme called GADPH Was used as a
50

level is RT-PCR (reverse transcriptase-polymerase chain reaction). Any measured changes in the parameters and meth
ods of measurement listed above can indicate an inhibitory or

stimulatory effect of the drug treatment

Although AZT has been a someWhat effective anti-HIV drug used in the clinical treatment of AIDS patients, it has become limited because of the development of resistance to this drug by the virus. Researchers WorldWide have been trying to ?nd neW drugs to counter this resistance problem. Here, it has been shoWn that the opioid antagonist naloxone
can inhibit HIV replication. It Will be seen that the objects set forth above, and those

control. No effect of drug treatment on expression of this enZyme Was noted indicating that the RT used in the assay condition Was not affected by the drug. That is GADPH can be considered a housekeeping gene of the T cell Which should have the same level of expression under all the drug
treatment conditions. The lack of an effect on GADPH also

55

proves that there Was no residual drug (naloxone or AZT)

present in the RNA sample used for analysis. The primer

sequence used for GADPH Were as folloWs: sense 5 prime

made apparent from the foregoing description, are ef?ciently

60

GGTGAAGGTCGGAGTCAACGGA-3 prime (SEQ ID NO:3); antisense 5 prime-GAGGGATCTCGCTCCTGG

GAAGA-3 prime (SEQ ID NO:4). The absence of or a decreased intensity of the gp41 band from infected cells treated With the drugs compared to control (non-treated) is
indicative of an inhibitory effect of the drug treatment on HIV. FIG. 4 is a RT-PCR analysis of RNA from HIV infected target cells Which had been treated With different molar con centrations of naloxone. The expression of the critical coat

attained and since certain changes may be made in the above construction Without departing from the scope of the inven tion, it is intended that all matters contained in the foregoing description or shoWn in the accompanying draWings shall be
interpreted as illustrative and not in a limiting sense. It is also to be understood that the folloWing claims are

65

intended to cover all of the generic and speci?c features of the invention herein described, and all statements of the scope of the invention Which, as a matter of language, might be the to fall therebetWeen. NoW that the invention has been described,

US 8,067,430 B1

SEQUENCE LISTING

<l60> NUMBER OF SEQ ID NOS:

<2ll> LENGTH: 21 <2l2> TYPE: DNA

<2l3> ORGANISM: artificial sequence

<220> FEATURE:

<223> OTHER INFORMATION: Sense Primer Sequence for Assay

<400> SEQUENCE: l

cagcgatagg agctctgttc c

21

<2ll> LENGTH: 21 <2l2> TYPE: DNA

<2l3> ORGANISM: artificial sequence

<220> FEATURE:

<223> OTHER INFORMATION: Antisense primer sequence for assay

<400> SEQUENCE: 2
gaccatttgc cacccatctt a 21

<2ll> LENGTH: 22 <2l2> TYPE: DNA

<2l3> ORGANISM: artificial sequence

<220> FEATURE: <223> OTHER INFORMATION:

Sense Primer sequence for GADPH

<400> SEQUENCE: 3

ggtgaaggtc ggagtcaacg ga

22

<2ll> LENGTH:

23

<2l3> ORGANISM: artificial sequence

<220> FEATURE:

<223> OTHER INFORMATION: Antisense Primer Sequence for GADPH

<400> SEQUENCE: 4

gagggatctc gctcctggga aga

23

45

What is claimed is: 1- A method Ofinhibiting reverse transcriptase activity in a

contacting the Virus With naloxone hydrochloride wherein the coat protein of the Virus is chosen from the group consisting

comprising the step of contacting the infected cell v(vith 31

solution consisting of naloxone hydrochloride having a molar 50
concentration between 10-4 M and 10-1 1 M.

cell infected With the human immunode?cienc Virus HIV

of gp41 and gpmo'

2. A method of modulating the expression of a coat protein of a human immunode?ciency Virus comprising the step of

US007879870B2

(12) Ulllted States Patent

Smith et a].
(54) TREATMENT OF INFLAMMATORY AND ULCERATIVE DISEASES OF THE BOWEL WITH OPIOID ANTAGONISTS

(10) Patent N0.:

(45) Date of Patent:
6,458,795 B1 6,664,270 B2
6,734,188 B1 6,737,400 B2

US 7,879,870 B2
Feb. 1, 2011

10/2002 Bergeron, Jr. 12/2003 Bergeron, Jr.

5/2004 Rhodes et al. 5/2004 Crain et al.

(76) Inventors: Jill P. Smith, 129 N. 30111 St, Camp Hill,

Ct., PA (US) Hummelstown, 17011; IanPA s. Zagon, (US) 17036; 589 Cook
Moshe Rogosnitzky, P. O. Box 386, TelZ
Stone IL (Us) 90840
* . _ . . . . 2008.

g;
Zoos/6261329 A1 11/2005

gages)? J
'

OTHER PUBLICATIONS
International Search Report for PCT/US2007/009228, mailed Jun. 3,

( ) Nonce'

SubJeCt.tO any ((111S(C11a1me5.thermft%1

Patent 15 exten e or a Juste un er

www.lowdosenaltreXone.com, as accessed on Mar. 21, 2007.

Smith et al., Low-Dose NaltreXone Therapy Improves Active

U~S~C- 15403) by 344 day5~

(21) Appl- NOJ 11/735,548

Crohns Disease, Am. J. Gastroenterol. 102, 1-9 (2007).

Penn State College of Medicine News Release, Penn State Research

(22) Filed:
(65)

Apr. 16, 2007

Shows Withdrawal Drug Offers Symptom Relief to Crohns Suffer ers (May 23, 2006). LDNiLow Dose NaltreXone, www.digitalnaturopath.com (as
accessed on Apr. 17, 2007). Low Dose NaltreXone: Informal Clinical Study Report, Jul. 1, 2005, www.stronadZieci.org (as accessed on Apr. 17, 2007).

Prior Publication Data Us Zoos/0015211 A1 J an. 17 , 2008

Primary ExamineriPhyllis G. Spivack

Related US. Application Data (74) Attorney, Agent, or FirmiLaurence Weinberger

(60) I1go\i8(i)o6nal application No. 60/745,119, ?led on Apr.

(51) Int Cl A61K 31/485

(57)

ABSTRACT

(200601)

Methods for the treatment of in?ammatory and ulcerative diseases of the bowel (e.g., Crohns disease and ulcerative colitis) with a therapeutically effective dose less than 50 mg.
of antagonists (e_g_, naltrexones nalmefene Or nalOX one) are disclosed. An embodiment of the invention includes

(58)

U-S- Cl- .................................................... .. Field of Classi?cation Search ..................... .. None

See application ?le for complete search history.

(56) References Cited
US. PATENT DOCUMENTS
4,987,136 A 4,994,466 A 5,013,739 A 1/1991 Kreek et al. 2/1991 Sherman et al. 5/1991 Bihari et al.

a method of pharmaceutical treatment comprising orally

admrmsterrng to a human subject having Crohn 5 disease or

ulcerative colitis a therapeutic pharmaceutical composition

once per day in the evening or at bedtime, wherein the phar maceutical composition comprises form about 3 mg to about 4.5 mg of naltrexone, nalmefene, naloxone, or a hydrochlo ride salt thereof in an immediate release solid dosage formu lation.

5,631,263 A
6,194,382 B1 6,395,705 B2

5/1997 Portoghese et a1.

2/2001 Crain et a1. 5/2002 Crain et a1.

20 Claims, 10 Drawing Sheets

US. Patent

Feb. 1, 2011

Sheet 1 0f 10

US 7,879,870 B2

BL4WWW
M**

L DN

ohmz?nv

m 1% %
12 16

Week

FIG. 1

US. Patent

Feb. 1, 2011

Sheet 2 0f 10

US 7,879,870 B2

A Response
LDN
100 _

Z! 4-WkS p0St_LDN

80

3,:x23.1

o 4 60

0 D

16

Week

B. Remission
60

mw?mwl Q0

2 3 4 1
0

12

%/ /
16

Week

FIG. 2

US. Patent

Feb. 1, 2011

Sheet 3 0f 10

US 7,879,870 B2

III!

'

_
EZZZZ
200 -

s post-LDN

****
* * >1<

****

150 -

QISuecsBtoirDnea
%

\ ;\ e
Week

FIG. 3

US. Patent

Feb. 1, 2011

Sheet 4 0f 10
II! Baseline
_ L DN

US 7,879,870 B2

m 4 -wks

posl-LDN
Bodily Pain

Role - Physical

Energy

General Health Perception

Physical Functioning

Social Functioning

Role - Emotional

Emotional Health

Week

FIG. 4

US. Patent

Feb. 1, 2011

Sheet 5 0f 10

US 7,879,870 B2

110

N0 DSS
105 -

100

LIJ U)
+'

95 +
I I I I I l | I I l

g
E a

1
D88: V 0

.97:

g2
g

D .97

g 0, E
100 _ . . .

D88
. . . . . .. . . . . . . . . . . . . . . . . . . .. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . , . ..

Tn
In'_

E cu
a)

95 -

Q:
90 -

85

80 _

9 Saline

$ 6.3 ug/kg NTX + 350 ug/kg NTX

75 I l I I l I l I I I

Day

US. Patent
40

Feb. 1, 2011

Sheet 6 0f 10

US 7,879,870 B2

1':

~k

**

u0

ST(9, MIFno:eEtaoMk:ndle)

MO

Saline

6.3
NTX

350
NTX

Saline

6.3
NTX

350
NTX

ug/kg ug/kg
No D88

ug/kg ug/kg
D88

120

100 -

1.

**

**

on O

ITSM(Wn1; oaemtEalMknre,)

(D O

NO

Saline

6.3
NTX

350
NTX

Saline

6.3
NTX

350
NTX

ug/kg ug/kg
No D88

ug/kg ug/kg
D88

FIG. 5C

US. Patent

Feb. 1, 2011

Sheet 7 0f 10

US 7,879,870 B2

N0 DSS
1.0
9 Saline

0.5

0.0 -

DSS: V
2 5.

// Treatment: ////////// / //// ///A

Necropsy: 0

DSS
2.0

1.5

1.0

0.5 -

0.0

Day

FIG. 6

US. Patent

Feb. 1, 2011

Sheet 8 0f 10

US 7,879,870 B2

30
**

2 1

**

0 5

_ _ _

Ec9o=20m wE

3mm 58% H

ll
Saline 6.3
NTX

350 NTX

Saline

6.3 NTX DSS

350

ug/kg ug/kg
N0 DSS

ug/kg ug/kg
NTX

FIG. 7A

U S. Patent

Feb. 1, 2011

Sheet 9 0110

US 7,879,870 B2

FIG. 7B

US. Patent

Feb. 1, 2011

Sheet 10 0110

US 7,879,870 B2

10

a
E 2(U
3

a 8
"E
(U
3 6*

]L

o
(D

o
(D

.2

.z

4,
2_

E 1(D

(D

n:

n:

Q_

Actin

n.

0
90 80

QReulantivey

60

50
40

30
20

- No D88 + Saline l:l D88 + Saline W D88 + 6.3 ug/kg NTX D88 + 350 ug/kg NTX

10*

STATS

FIG. 8

US 7,879,870 B2
1
TREATMENT OF INFLAMMATORY AND ULCERATIVE DISEASES OF THE BOWEL WITH OPIOID ANTAGONISTS CROSS REFERENCE TO RELATED APPLICATIONS

2
Treatment of Crohns disease usually includes administra

tion of anti-in?ammatory drugs, including compounds

designed to reduce the in?ammatory response, such as corti

costeroids, cyclosporine, and azathioprine, Which often lead

to serious side effects. Major advances in the understanding of the pathogenesis of IBD have led to the development of novel immunotherapies. Such treatments include the admin istration of chimeric monoclonal antibodies speci?c for mol ecules expressed by the T-cells population or antibodies spe ci?c for cytokines knoWn to be central to the pathogenesis of

This application claims the priority of US. provisional patent application No. 60/745,119, ?led on Apr. 19, 2006, incorporated herein by reference in its entirety.
FIELD

mucosal in?ammation (anti-tumor necrosis factor, TNF). Although this speci?c immunotherapy has helped those With
Crohns disease, still about 20% do not respond to this treat ment and many cannot continue this therapy due to untoWard side effects. Additionally, treatment With the monoclonal

Example embodiments of the invention include methods for the treatment of in?ammatory and ulcerative diseases of the boWel (e.g., Crohns disease and ulcerative colitis) With

loW dose opioid antagonists (e.g., naltrexone, nalmefene or naloxone), pharmaceutical compositions for use in such methods, and methods for the manufacture of such pharma ceutical compositions.
BACKGROUND

antibody in?iximab (sold as REMICADE, a registered trademark of Centocor, Inc. of Malvern, Pa.) is expensive
With each infusion costing in excess of US$3,000. Ulcerative colitis is a form of colitis, an in?ammatory disease of the intestine, usually the colon, that includes char acteristic ulcers. Symptoms of active disease usually include

20

Chronic relapsing and remitting in?ammation of the gas

trointestinal tract are hallmarks of ulcerative colitis and
25

diarrhea mixed With blood, usually accompanied With vary ing degrees of abdominal pain, from mild discomfort to severely painful cramps. Although ulcerative colitis has no
knoWn cause, there is a presumed genetic component to sus

Crohns disease, conditions termed in?ammatory boWel dis

eases (IBD). The peak age of onset of this disease is betWeen the ?rst and fourth decades of life, With a prevalence of 100-200 per 100,000 in Europe and North America.

ceptibility. The disease may be triggered in a susceptible

person by environmental factors. Although dietary modi?ca

tion may reduce the discomfort of a person With the disease,

ulcerative colitis is not thought to be caused by dietary fac

30

In?ammatory boWel disease accounts for signi?cant morbid

tors. As With Crohns disease, treatment includes administra

ity and decreased quality of life, and is responsible for nearly

US$20 billion in annual medical costs in the United States.

tion of anti-in?ammatory drugs, immunosuppression, and

biological therapy targeting speci?c components of the

immune response.

Although there has been progress in de?ning the pathogen

esis of these diseases, their cause remains obscure. The cur rent most comprehensive hypothesis is that IBD is a hetero geneous group of diseases that have a ?nal manifestation,
35

When anti-in?ammatory therapies fail, colectomy is occa

sionally necessary, Which is considered to be curative for

ulcerative colitis. Surgery is generally reserved for complica

tions of Crohns disease or When disease that resists treatment With drugs is con?ned to one location that can be removed.
40

Which is mucosal in?ammation, and that several genetic and environmental factors are implicated in the pathogenesis of
the disease. Because of the name, IBD is often confused With irritable boWel syndrome (IBS), a troublesome but much less seri ous condition. Irritable boWel syndrome is a gastrointestinal disorder characterized by altered boWel habits and abdominal

Surgery is also used to manage complications of Crohns disease, such as ?stulae and small boWel obstructions, and for

resection and anastomosis (e.g., ileocolonic resection). Sur

gery rarely cures Crohn s disease, and recurrence often reap pears in previously unaffected areas of the intestine.
45

pain in the absence of detectable structural abnormalities,

such as in?ammation or ulceration. Irritable boWel syndrome is not an in?ammatory disease and it is not characterized by

Accordingly, a continuing need exists for effective phar macological treatments of in?ammatory boWel disease, such
as Crohns disease and ulcerative colitis.

ulceration of the boWel. No clear diagnostic markers exist and no pathognomonic abnormalities have been identi?ed for
IBS, so all de?nitions of the disease are based on the clinical

SUMMARY
50

presentation. Until recently, many physicians did not con

sider IBS to be a disease at all; they vieWed it as nothing more

Example embodiments of the invention include methods for the treatment of in?ammatory and ulcerative diseases of

than a somatic manifestation of psychological stress. Typical

chemotherapies for IBS include stool bulking agents, antis pasmodics, and antidiarrheal agents.
In contrast, Crohns disease is an in?ammatory disease
55

the boWel (e.g., Crohns disease and ulcerative colitis) With loW dose opioid antagonists (e.g., naltrexone, nalmefene or naloxone), pharmaceutical compositions for use in such methods, and methods for the manufacture of such pharma

characterized by transmural, patchy, granulomatous in?am

mation of any part of the gastrointestinal tract, although it is
common in the ileocecal area. Symptoms of Crohns disease

ceutical compositions.
In another example embodiment, the invention includes the
use of an opioid antagonist in the preparation of a loW dose pharmaceutical composition for the treatment of a boWel disease characterized by in?ammation or ulceration of the intestinal Wall. In yet another embodiment, the invention includes a method for treating a boWel disease comprising administer ing to a subject in need thereof a loW dose of an opioid antagonist effective to treat a boWel disease in the subject

include abdominal pain, diarrhea, gastrointestinal bleeding, malabsorption, and Weight loss. Although the etiology is
unknoWn, research suggests it involves a complex interplay of

60

environmental, genetic, microbial, immune, and nonimmune

factors. Biopsies obtained from the boWel in subjects With Crohns disease reveal in?ammatory cells suggesting that the
boWel is either reacting immunologically to a stimulus or the
65

endogenous immune system of the gastrointestinal track is off

balance.

(e. g., a human), Wherein the boWel disease is characterized by

in?ammation or ulceration of the intestinal Wall.

US 7,879,870 B2
3
In still yet another embodiment, the invention includes a pharmaceutical composition for use in the treatment of a boWel disease characterized by in?ammation or ulceration of

4
preferred embodiments, taken in conjunction With the accom

panying draWings that illustrate, by Way of example, prin

ciples of the invention.
BRIEF DESCRIPTION OF THE DRAWINGS

the intestinal Wall, the pharmaceutical composition compris

ing a loW dose (e.g., less than about 5 mg) of an opioid

antagonist.
The invention provides heretofore unknown evidence that loW dose naltrexone, an opioid antagonist, is a safe and effec
tive treatment for Crohns disease. In summary, an open

FIG. 1 illustrates mean Crohns Disease Activity Index scores associated With LDN therapy in human patients. FIG. 2 illustrates the percent of patients responding, and

the percent of patients achieving remission, to LDN therapy.

FIG. 3 illustrates mean In?ammatory BoWel Disease Ques tionnaire scoresiSEM associated With LDN therapy in

labeled prospective trial of human subjects Was conducted, as described more fully herein beloW. The safety and e?icacy of loW dose naltrexone (LDN) Was tested in patients With

human patients.
FIG. 4 illustrates mean SF-36 Health Survey scoresiSEM

active Crohns disease. Eligible subjects With histologically

and endoscopically con?rmed active Crohns having a Crohns Disease Activity Index (CDAI) score of 220-450 Were enrolled in a study using 4.5 mg naltrexone/day. In?ix
imab Was not alloWed for a minimum of eight Weeks prior to

associated With LDN therapy in human patients.

FIG. 5A illustrates the mean Weight of laboratory animals

receiving either normal drinking Water (upper panel) or 2% DSS in drinking Water (loWer panel). FIG. 5B illustrates the total food intake per animal during the DSS/NTX experiment.
20

study initiation or While in the study. Other therapy for

Crohns disease that Was at a stable dose for four Weeks prior
to enrollment Was continued at the same doses. Patients com

FIG. 5C illustrates the total Water intake per animal over the course of the study.

pleted the In?ammatory BoWel Disease Questionnaire

(IBDQ) and the Short-Form (SF-36) quality of life sur
veys and CDAI scores Were assessed pretreatment, every four
25

FIG. 6 illustrates the reduction of daily modi?ed Disease Activity Index scores in laboratory animals treated With nal
trexone.

FIG. 7A illustrates the difference in the in?ammation of

Weeks on therapy, and four Weeks after completion of the

study drug. LDN Was administered by mouth each evening

for a tWelve-Week period. Seventeen patients With a mean
CDAI score of 356127 Were enrolled. CDAI scores decreased 30

longitudinal sections of the distal colon upon staining and evaluation. FIG. 7B illustrates representative stained sections
of distal colon. FIG. 8 illustrates the real-time RT-PCR quanti?cation of total RNA from the distal colons of control mice, mice With untreated colitis, and mice With colitis treated With naltrex
one.

signi?cantly (p:0.0l) With LDN, and remained loWer than

baseline four Weeks after completing therapy. Eighty-nine

percent of patients exhibited a response to therapy and 67% achieved a remission (p<0.00l). Improvement Was recorded in both quality of life surveys With LDN compared to base line. No laboratory abnormalities Were noted. The invention also provides heretofore unknown evidence
35

DETAILED DESCRIPTION

Example embodiments of the invention include methods for the treatment of in?ammatory and ulcerative diseases of

that the opioid antagonist naltrexone reduces in?ammation of

the boWel in a chemically-induced mouse model of ulcerative

the boWel With loW dose opioid antagonists (e.g., less than
about 5 mg naltrexone, nalmefene or naloxone per day),
40

colitis, as described more completely herein beloW. Brie?y, laboratory mice received either untreated drinking Water or Water containing 2% dextran sulfate sodium (DSS) for six days. Three days after DSS introduction, animals Were administered 0.1 mL saline (control) or naltrexone (NTX,

pharmaceutical compositions for use in such methods, and methods for the manufacture of such pharmaceutical compo
sitions. In an embodiment, the invention relates to the treatment of in?ammatory boWel disease, such as Crohns disease and ulcerative colitis. In?ammatory boWel disease (IBD) is a

45

6.3 or 350 ug/kg) daily for six days. Disease activity index
(DAI) scores Were calculated daily. Mice Were necropsied on day nine and colon in?ammation Was analyZed histologi

group of in?ammatory conditions of the large intestine and

the small intestine. Although similar in name, it should not be confused With irritable boWel syndrome (IBS), Which is a different condition having a different pathology and different symptoms. Principal forms of IBD are Crohns disease and

cally. Colonic RNA Was evaluated by microarray and real time RT-PCR. By day 4, DSS-treated animals had signi?cant
Weight loss (p:0.006) and higher DAI scores (p<0.00l) com
pared to Water controls. DSS-treated mice that received nal

50

ulcerative colitis (UC), although other forms include col

trexone (350 ug/kg) had less Weight loss, loWer DAI scores,
and less in?ammation compared to DSS mice treated With

lagenous colitis, lymphocytic colitis, ischemic colitis, diver sion colitis, Behcets syndrome, infective colitis, and indeter
minate colitis. Crohns disease and UC differ is the location
55

saline. RNA encoding pro-in?ammatory cytokines IL-6 and

IL-l2 Was signi?cantly elevated in DSS-treated mice, and
after naltrexone treatment these cytokine RNAs Were decreased to levels near or equal to that in mice Without

and nature of the in?ammatory changes in the intestine. Crohn s may affect any part of the gastrointestinal tract, from
mouth to anus, although the terminal ileum is the area most

colitis. Expression of the transcription factors STAT3 and STAT4, doWnstream effectors of cytokine signaling, Was also up-regulated in DSS-treated mice and similarly reversed by
naltrexone. Naltrexone therefore appears to have reversed

commonly involved. Ulcerative colitis, in contrast, is gener ally restricted to the colon and the rectum. Microscopically, ulcerative colitis is restricted to the mucosa (i.e., the epithelial lining), While Crohn s disease may affect the entire thickness of the intestinal Wall. Finally, Crohns disease and ulcerative
colitis present With extra-intestinal manifestations such as

disease manifestations and histologic evidence of in?amma tion in DSS-induced colitis. Other features and advantages of the present invention Will be apparent from the folloWing more detailed description of

liver problems, arthritis, skin manifestations and eye prob

65

lems. In?ammatory boWel disease may present With any of the

folloWing symptoms: abdominal pain or discomfort, abnor

US 7,879,870 B2
5
mal bowel movement frequency (e.g., diarrhea), intestinal
stenosis or ?stulization, perianal discomfort or pruritis, vom

6
salt thereof (e.g., naltrexone hydrochloride, nalmefene hydrochloride, and naloxone hydrochloride).
According to an embodiment of the invention, the opioid
antagonist is administered as a loW dose. For example, the

iting, hematochezia (i.e., bloody stool), Weight loss and vari

ous associated complaints or diseases. IBD may also present With various ?stulae, including one or more gastrocolic ?s

loW dose of the opioid antagonist may be from about 1.75 mg

to about 4.5 mg, or alternatively form about 1.75 mg to about 3 mg, or in another embodiment from about 3 mg to about 4.5

tulae, gastrojejunocolic ?stulae, enterocutaneous ?stulae, anorectal ?stulae, enterovaginal ?stulae, enterovesical ?stu lae, and enteroenteral ?stulae. Diagnosis of in?ammatory boWel disease is generally by colonoscopy With biopsy of pathological lesions or by radiographic (e.g., X-ray) exami
nation, Which often reveals in?ammation or ulceration of the

mg. The loW dose of opioid antagonist may be administered

once per day, eg in the evening or at bedtime so that tmax occurs shortly after sleep commences. Accordingly, in an

embodiment, the opioid antagonist is provided as a pharma

ceutical composition, Which may be administered once per day in the evening or at bedtime. In another embodiment, the

ileum or colon (e.g., the ileocecum).

Accordingly, an embodiment of the invention includes a

method for treating a boWel disease comprising administer

ing to a subject in need thereof a loW dose of an opioid antagonist effective to treat a boWel disease in a subject (e.g., a human patient), Wherein the boWel disease is characterized by in?ammation or ulceration of the intestinal Wall or any

pharmaceutical composition is administered once per day in the morning or after Waking from sleep.
Another embodiment of the invention includes a method of

pharmaceutical treatment comprising orally administering to

a human subject having Crohn s disease or ulcerative colitis a

portion thereof, eg the mucosa. As used herein a boWel disease may be characterized by abdominal pain or discomfort, abnormal boWel movement frequency, intestinal stenosis or ?stulization, perianal dis comfort or pruritis, or abnormal stool consistency. Further
more, a boWel disease may be characterized by in?amma tion or ulceration of the small intestine (e.g., the ileum) or

therapeutic pharmaceutical composition once per day in the

20

evening or at bedtime (or once per day in the morning or after

Waking from sleep), Wherein the pharmaceutical composition

comprises form about 3 mg to about 4.5 mg of naltrexone, nalmefene, naloxone, or a hydrochloride salt thereof per dose, Which may be an immediate release solid dosage formulation. The invention also relates to pharmaceutical compositions

25

colon (e.g., the ileocecum). A boWel disease may also be characterized by the presence of a ?stula, eg of the gas trointestinal system. In particular, a boWel disease may include in?ammatory boWel disease (IBD), such as
Crohns disease or ulcerative colitis. A boWel disease as
30

and the preparation thereof. The pharmaceutical composition

may be formulated as a solid dosage form suitable for oral administration. Furthermore, the solid do sage form may be an

used herein does not include irritable boWel syndrome per se. Another embodiment of the invention includes the use of

immediate release formulation comprising an opioid antago nist and an excipient, Which may be selected from, eg the group consisting of sucrose, cellulose, and combinations

thereof. In another embodiment, the loW dose pharmaceutical

composition may be formulated as a liquid dosage form suit

an opioid antagonist in the preparation of a loW dose pharma

ceutical composition for the treatment of a boWel disease

(e. g., IBD) characterized by in?ammation or ulceration of the

intestinal Wall or any portion thereof, e. g. the mucosa. In still yet another embodiment, the invention relates to a pharmaceutical composition for use in the treatment of a

boWel disease characterized by in?ammation or ulceration of the intestinal Wall or any portion thereof, eg the mucosa, the pharmaceutical comprising a loW dose of an opioid antago nist. In still another embodiment, the invention relates to a phar maceutical composition for use in the treatment of a ?stula, eg a connection from the boWel to either the skin, another

40

45

part of the boWel, the bladder, or vagina, the pharmaceutical

comprising a loW dose of an opioid antagonist. The ?stula
may be incident, to or associated With, a boWel disease char acterized by in?ammation or ulceration of the intestinal Wall or any portion thereof, e. g. the mucosa. In another embodiment, the invention includes a method for treating a boWel disease comprising administering to a
50

able for oral administration. For example, the liquid dosage form may comprise an opioid antagonist and a liquid carrier, Which may comprise Water. Similarly, the pharmaceutical composition may be formulated as a topical dosage form suitable for topical administration. It is especially advantageous to formulate pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. A unit dosage form refers to physi cally discrete units suited as unitary dosages for the subjects to be treated; each unit containing a predetermined quantity of opioid antagonist calculated to produce the desired thera peutic effect in association With the required pharmaceutical vehicle. The speci?cation of unit dosage forms may be dic tated by and directly dependent on the unique characteristics of the particular opioid antagonist and the limitations inherent
in the art of compounding such a compound for the treatment ofa boWel disease.

Opioid antagonists may be formulated for administration using physiologically acceptable carriers or excipients in a
manner fully Within the skill of the art. For example, the
55

subject in need thereof a pharmaceutical composition com

prising a loW dose of an opioid antagonist effective to treat a

opioid antagonists, optionally With the addition of pharma ceutically acceptable excipients, may be suspended or dis
solved in an aqueous medium, With the resulting solution or

boWel disease in said subject, Wherein said boWel disease is characterized by in?ammation or ulceration of the intestinal Wall. The amount of opioid antagonist may be from about 1.75 mg to about 4.5 mg per dose, or alternatively form about
1.75 mg to about 3 mg per dose, or in another embodiment from about 3 mg to about 4.5 mg per dose.
60

suspension then being sterilized. An opioid antagonist may be

orally administered, for example, With an inert diluent or an

edible carrier. The opioid antagonist and other ingredients

may also be enclosed in a hard or soft shell gelatin capsule or

As used herein, an opioid antagonist may be selected from the group consisting of naltrexone, nalmefene, nalox

compressed into tablets. For oral therapeutic administration, the opioid antagonist may be incorporated With excipients
and used in the form of ingestible tablets, buccal tablets,

one, metabolites thereof having opioid antagonist activity,

troches, capsules, elixirs, suspensions, syrups, Wafers, and

65

pharmaceutically acceptable salts thereof, prodrugs thereof,

and combinations thereof. For example, an opioid antagonist may be naltrexone, nalmefene, naloxone, or a hydrochloride

the like.

For topical administration, the opioid antagonist may be provided in the form of ointments, creams, gels, lotions,

US 7,879,870 B2
7 shampoos, powders (including spray powders), pessaries,
tampons, sprays, dips, aerosols, or drops (e.g., eye ear or nose

8
that may be cleaved to reveal an intermediate compound that

subsequently decomposes to yield the active compound. The

prodrug moieties may be metaboliZed in vivo by esterases or

drops). The opioid antagonist may also be provided in the

form of a transdermal or percutaneous drug delivery system, eg a transdermal patch.

by other mechanisms to carboxylic acids. Examples of pro

drugs and their uses are well known in the art. Prodrugs of

The percentage of the opioid antagonist in the pharmaceu

tical compositions may, of course, be varied. The relative

opioid antagonists may be prepared in situ during their ?nal isolation and puri?cation or by separately reacting the puri
?ed compound in its free acid or base form with a suitable

amount of the opioid antagonist in such therapeutically useful

compositions is such that a suitable dosage will be obtained.

Furthermore, the pharmaceutical composition may be formu

lated as an immediate release formulation (preferred), an intermediate release formulation, or an extended release for

derivatiZing agent. See, eg R. B. Silverman, The Organic Chemistry of Drug Design and Drug Action, Academic Press, Chp. 8 (1992).
EXAMPLES

mulation according to methods well known in the art. Opioid antagonists are typically alkaloids that contain a
basic functional group, such as amino group, and are thus

Crohn s Disease

capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable acids. The term pharmaceuti
cally acceptable salts in this respect, refers to the relatively non-toxic, inorganic and organic acid addition salts of opioid antagonists. These salts may be prepared in situ during the ?nal isolation and puri?cation of the compounds, or by sepa rately reacting a puri?ed compound in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed. Representative salts include the hydrohalide (includ
20

In the following detailed example, low dose naltrexone, an

opioid antagonist, is shown to be a safe and effective treat ment for Crohns disease. In a human clinical experiment,

eligible patients were both male and female, at least eighteen years of age, and with the con?rmed diagnosis of Crohns

disease by either endocopic or radiographic procedures.

25

ing hydrobromide and hydrochloride), sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stear ate, laurate, benZoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, 2-hydroxyethylsulfonate, lau
rylsulphonate salts and the like. See, eg Berge, et al., Phar maceutical Salts, .1. Pharm. Sci. 66, 1-19 (1977).
In other cases, an opioid antagonist may contain one or more acidic functional groups and thus be capable of forming

Patients had moderate to severely active disease as de?ned by a Crohns Disease Activity Index (CDAI score) of >220 and <450. See, Best, et al., Development of a Crohns disease

activity index. National Cooperative Crohn s Disease Study,

Gaslroenlerology 70, 439-44 (1976). Patients taking stable doses of aminosalicylates, immunomodulators, corticoster
30

oids, or antibiotics were permitted to enter the study, and they were continued at the same dosage throughout the trial.

pharmaceutically acceptable salts with pharmaceutically acceptable bases. The term pharmaceutically acceptable
salts in these instances refers to the relatively non-toxic,

Women of childbearing age were permitted to enroll and, if not surgically sterile, were required to use adequate contra ception (de?ned as oral or depot contraceptive, IUD, or bar

rier plus spermicide) for the duration of the study. These

35

women were required to continue adequate contraception for

three months after the completion of the study. Exclusion

criteria included women who were pregnant or breastfeeding,

inorganic and organic base addition salts of opioid antago

nists. These salts can likewise be prepared in situ during the ?nal isolation and puri?cation of the compounds, or by sepa

rately reacting the puri?ed compound in its free acid form

with a suitable base, such as the hydroxide, carbonate or

40

subjects with an ileostomy, colostomy, ileorectal anastomo sis, or short bowel syndrome from surgery, and patients with abnormal liver function tests. Subjects taking tacrolimus, cyclosporine, mycophenolate, or in?iximab within eight
weeks of enrollment were excluded.

bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Rep resentative organic amines useful for the formation of base

TABLE 1
45

Patient Demographics
Mean age 1 SEM (years) 42.1 r 2.6

addition salts include ethylamine, diethylamine, ethylenedi amine, ethanolamine, diethanolamine, piperaZine and the like. See, Berge, Ibid. Pharmaceutically acceptable includes those compounds,
materials, compositions, or dosage forms that are, within the
scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without exces

(range)
Gender, N (% of patients)
50

(23-63)
3 (18%) 14 (82%)
72 r 4

Male Female
Mean body weight 1 SEM (kg)

(range)
Disease site
55

(53-101)
2 (12%) 10 (59%) 5 (29%)

sive toxicity, irritation, allergic response, or other problem or

complication, commensurate with a reasonable bene?t/risk ratio.

Small bowel Small bowel & colon Colon

The invention also pertains to prodrugs of opioid antago

nists. Prodrugs are compounds that are converted in vivo to active forms. Prodrugs can be used to alter the biodistribution or the pharmacokinetics of a particular compound. For example, a carboxylic acid group can be esteri?ed, eg with a methyl group or an ethyl group to yield an ester. When the ester is administered to a subject, the ester is cleaved, enZy

Past resection performed, N (% of patients) 8 (47%) Prior anti-TNF-ot therapy, n (% of patients) 13 (76%) Concomitant meds for Crohns, N (% of patients)

Arninosalicylates
Immunomodulators Glucocorticoids Antibiotics
65

11 (65%)
8 (47%) 4 (24%) 1 (6%)

matically or non-enZymatically, reductively, oxidatively, or

hydrolytically, to reveal the anionic group. An anionic group

may be esteri?ed with moieties (e.g., acyloxymethyl esters)

The study was an open-labeled pilot investigation to evalu ate safety, toxicity, and response to LDN in subjects with

US 7,879,870 B2
10
active Crohns disease. Eligibility Was assessed by telephone,
and potential candidates Were scheduled for a screening visit.
P-values for binary outcomes response and remission Were

calculated using the exact test for binomial proportions.

The characteristics of the patients at enrollment are shoWn

At the screening visit, patients Were subjected to a history and

physical examination and laboratory testing (chemistry panel, complete blood count [CBC], urinalysis, and eryth
rocyte sedimentation rate [ESR]). Patients Were dispensed a seven-day diary to record symptoms of frequency of diar

in Table 1, including age, gender, and body Weight. Most

patients had both small boWel and colonic disease, and tWo

patients had active perianal ?stulas. Eight patients had prior

surgical resection performed for their Crohns disease. Sev enty-six percent of patients had prior treatment With anti
TNF-ot therapy and Were either allergic, intolerant, or unre sponsive to this medication. Concomitant medications for

rhea, abdominal pain, and general Well-being. Within four

teen days, patients returned for assessment and calculation of the CDAI score. Qualifying subjects Were dispensed medica tion and given a neW diary in order to calculate the sub sequent months CDAI score at the conclusion of this visit (baseline).
Patients returned after tWo Weeks for an interim visit to evalu ate side effects and perform a CBC. FolloW-up visits Were

Crohns disease taken by patients throughout the study are

also shoWn in Table 1 hereinabove. Statistical analysis shoWed that there Was no signi?cant

change in body Weight from screening visit through Week 16

of the study (data not shoWn). TWo patients elected to discon tinue taking routine medications for Crohns disease prior to
Week 12 and symptoms of Crohns disease recurred in one of them. Data from both patients Were analyZed With an intent to -treat paradigm. The tWo subjects With entercutaneous and
20

scheduled for Weeks 4, 8, 12, and 16. Naltrexone hydrochloride Was compounded into capsules containing 4.5 mg by GMP-approved standards at a phar macy. Because the dosage used in this study Was loWer than
the current FDA-approved dose of 50 mg, it is referred to as loW dose naltrexone or LDN. Quality assurance of packag

rectovaginal ?stulas had closure of the ?stulas With LDN

ing and purity Were con?rmed an analytical research labora tory. Patients Were treated With LDN orally each evening for three months.A monthly supply of medication Was dispensed to patients. On the ?rst visit, an additional ten-day supply of LDN Was provided in the event of an appointment delay. Subjects Were required to bring the vials to each appointment

therapy.
Referring to the draWings, FIG. 1 illustrates the mean

Crohns Disease Activity Index (CDAI) scoresiSEM, shoWn at baseline (Week 0), Weeks 4, 8, and 12 after initiation of
25

LDN therapy and 4 Weeks after discontinuation of LDN

therapy (Week 16). ****:Signi?cantly different from base

line at p<0.0001. FIG. 2 shoWs the percent of patients

for counting and drug accountability; extra capsules Were

returned on the day of the visit and another months supply

responding (FIG. 2A), and the percent of patients achieving

remission (FIG. 2B), to LDN therapy, shoWn at Weeks 4, 8,
30

dispensed.
Routine blood Work including CBC, chemistry panel, and
ESR Were assessed monthly. In addition, urine tests and preg nancy tests Were done for monitoring and safety purposes

and 12 and 4 Weeks after discontinuation of LDN therapy (Week 16). FIG. 3 illustrates mean In?ammatory BoWel Dis

pretreatment and at each monthly visit. C-reactive protein

(C-RP) Was measured at baseline and at Week 12. [Met5]

ease Questionnaire (IBDQ) scoresiSEM, shoWn at baseline (Week 0), Weeks 4, 8, and 12 after initiation of LDN therapy, and 4 Weeks after discontinuation of treatment (Week 16).
35

enkephalin levels Were determined by radioimmunoassay (RIA) at baseline and Weeks 4, 8, 12, and 16.
In order to assess the effect of LDN on disease activity,

Signi?cant differences from baseline are shoWn as * *p<0.01,

patients kept a Crohns symptom diary for the seven days

preceding each visit for calculation of the CDAI score. To

***p<0.001, and ****p<0.0001. In FIG. 4, mean SF-36 Health Survey scoresiSEM are shoWn at baseline (Week 0), Weeks 4, 8, 12 of LDN therapy, and 4 Weeks after discontinu ation of treatment (Week 16) for each of the parameters mea
40

assess quality of life, patients completed tWo standardized quality of life surveys, the In?ammatory BoWel Disease Questionnaire (IBDQ) and SF-36 Health Survey. See, Irv
ine, et al., Quality of life: a valid and reliable measure of

sured by the SF-36 Health Survey. Signi?cant differences from baseline values included the folloWing: *p<0.05,

**p<0.01, ***p<0.001, and ****p<0.0001.

CDAI scores Were used to measure the patients disease
45

therapeutic e?icacy in the treatment of in?ammatory boWel disease. Canadian Crohns Relapse Prevention Trial Study

activity and in?ammatory response to LDN therapy. Mean CDAI scores (FIG. 1) at Weeks 4, 8, and 12 folloWing the
initiation of LDN therapy Were 41%, 55% and 49%, respec tively decreased from baseline. Four Weeks after discontinu ation of therapy (Week 16), the mean CDAI score Was 45%

Group, Gaslroenlerology 106(2), 287-96 (1994); BraZier, et

al., Validating the SF-36 health survey questionnaire: neW outcome measure for primary care, BMJ305 (6846), 160-04

(1992).
The safety and toxicity of LDN Were assessed by adverse

50

less than baseline and not statistically different from the mean scores measured during the therapy. FIG. 2 shoWs the per

events, laboratory parameters, and vital signs. Nonhemato logic and hematologic toxicity Were determined by the WHO criteria. See, Oken, et al., Toxicity and response criteria of the Eastern Cooperative Oncology Group, Am. J. Clin. Oncol. 5(6), 649-55 (1982).
An intent-to-treat analysis Was performed in Which the available data from all evaluable patients Were included in the

centage of patients responding to therapy (FIG. 2A), as Well

as the percentage of subj ects achieving a remission of disease (FIG. 2B). At one month after treatment, 76% had achieved a response to therapy (a decrease in the CDAI score by 70 points), and at 8 and 12 Weeks, 88% shoWed a response. Four Weeks after discontinuation of LDN, 73% continued to shoW a response. At one month after starting LDN therapy, 29% of the patients had achieved a remission (a CDAI score of 150 points or less), and at Weeks 8 and 12 of LDN therapy, 53%

55

statistical analysis. The parameters of measurement (CDAI scores, laboratory values, and quality of life surveys) Were analyZed by a biostatistician comparing baseline values to those obtained monthly and four Weeks post-therapy. A lon gitudinal data analysis based upon the linear mixed-effects model Was applied using PROC MIXED program (available from SAS Institute Inc., Cary, NC.) The Bonferroni statis tical method Was used to adjust signi?cance Where analysis including multiple comparisons to the baseline Were made.

60

and 47%, respectively, had achieved remission (FIG. 2B).

Four Weeks after discontinuation of LDN therapy, 33% of the subjects Were in clinical remission. Therefore, at some point during the 16-Week trial, 89% of patients exhibited a response (p<0.001), and of 67% achieved a remission (p:0.07) With

65

LDN.

TWo standardized quality of life surveys, the In?ammatory

BoWel Disease Questionnaire (IBDQ) (FIG. 3) and the SF-36

US 7,879,870 B2
11
Health Survey (FIG. 4) Were administered to patients receiv ing LDN treatment. By both measures, patients experienced a signi?cant improvement in their quality of life on LDN

12
indicated that for tWo separate quality of life surveys a sig ni?cant difference from baseline occurred in those individu
als on LDN. Moreover, even one month after discontinuation

therapy. With regard to the IBDQ survey, signi?cant improve

ment in quality of life Was noted compared to baseline at Weeks 4, 8, and 12 on LDN, as Well as one month after

completion of treatment.

of LDN therapy, the quality of life remained better in almost all parameters measured for these patients. As of the date of application for patent hereof, a double-blinded placebo-con trolled Phase II clinical trial of human subjects has not
revealed any serious adverse events. Treatment With LDN provides advantages over other stan

Patients experienced a signi?cant improvement in quality

of life in a variety of parameters as measured by the SF-36

Health Survey (FIGS. 4A-4H). At Weeks 4, 8, and 12 of

therapy With LDN resulted in a 5- to 8-fold improvement in physical role scores (FIG. 4A) and a 61-65% improvement in

dard therapy for Crohns disease. The safety pro?le of LDN appears to be excellent in this study, With infrequent and
minor side effects and no knoWn suppression of immunity or

bodily pain (FIG. 4B). Energy scores at Weeks 8 and 12 of

LDN treatment (FIG. 4C) Were at least 2-fold greater than at the time of initiation of therapy, Whereas the scores for health

increased risk of secondary infections. Cortiocosteroids have

signi?cant long-term risks of Weight gain, osteoporosis, cata

racts, and glucose intolerance. The immunomodulators meth

perception (FIG. 4D) Were 33% and 49%, respectively,

greater than baseline. At 4 and 8 Weeks of LDN therapy, the

otrexate, aZathioprine, 6-mercaptopurine, and cyclosporine

all impair immune function increasing the risk for infections and perhaps malignancies. In addition to immunosuppres sion
20

physical function (FIG. 4E) Was 23% greater than baseline values. Social function (FIG. 4F) Was 70% greater than base line at Weeks 4, 8, and 12, but Was only statistically different at Week 8. Role-emotional (FIG. 4G) and emotional health (FIG. 4H) Were comparable to baseline values at Weeks 4, 8,
and 12 of LDN treatment. At 4 Weeks after termination of

With the neW anti-TNF-ot compounds, these drugs can also increase the risk of reactivation of tuberculosis and induce a

lupus-like reaction, serum sickness syndrome, or anaphy laxis. Some investigators have suggested that in?iximab (an
anti-TNF-ot compound) or other immunomodulators may

LDN (i.e., Week 16), all parameters except emotional health

shoWed improvement ranging from 27% to an 8-fold improvement over baseline. At Weeks 4, 8, and 12 of LDN therapy there Was no change from baseline in CBC or chemistry values. Liver panels Were not altered from baseline levels at Week 12. Assessment of CBC at Week 2 of LDN therapy Was comparable to baseline values. C-reactive protein levels decreased from a median
25

increase the risk of malignancies, in particular lymphomas;

hoWever, it is unclear Whether the risk is due to the disease itself rather than the medication. Higher doses of naltrexone

(i.e., 50 mg) used for alcohol and opioid abuse have been
30

reported to elevate liver transaminases. In contrast, the use of LDN herein at 4.5 mg daily did not change liver transami nases during treatment. In?iximab has become a standard medical therapy for

value of 2.6 (normal <0.8) at baseline to a value of 0.9 by the 12 Week of treatment, and this change Was statistically sig
ni?cant (p:0.03). The ESR decreased from a mean baseline value of 23310.4 mm/hr to 17.9103 mm/hr, Which Was also
35

patients With ?stulizing disease associated With Crohns dis

ease. It is of interest that tWo subjects in our study With enterocutaneous ?stulas noted closure With LDN When they

had not previously responded to in?iximab. Perhaps closures

of the ?stulas may be related to decreased intestinal secre tions or mucosal healing. Perhaps the ?stulas closed as a result of decreased number of boWel movements and

signi?cant (p:0.04). Baseline plasma enkephalin levels Were

9512.8 pg/mL, and decreased to a value of3.6:1.0 pg/mL at

Week 12 of LDN therapy, but this difference in plasma enkephalin levels Was not statistically signi?cant. The mo st frequently reported side effect With LDN therapy
Was sleep disturbances, and this Was noted in 7 patients; one

improved mucosal ?uid absorption.

40

Medical care for IBD is costly. Aminosalicylate therapy

can cost several hundred dollars per month, and an in?iximab

reported unusual dreams. Five subjects changed the timing of

LDN from the evening to morning due to insomnia. In no instance Was a dose reduction necessary for sleep distur

bances. Other rarely reported events included nausea (NIl ),

45

hair thinning (NIl ), blurred vision (NIl), irritability (NIl ),

mood sWings (NIl), and mild disorientation (NIl).
The results of this pilot study are the ?rst to shoW that LDN

infusion generally exceeds several thousand dollars (not men tion of the time aWay from the Workplace for intravenous administration). Naltrexone is a generic medication and the cost is therefore inexpensive. Moreover, effective mesalamine (PENTASA, a trademark of Ferring B. V. of The Netherlands) therapy may require up to 8-16 tablets per

therapy signi?cantly decreases symptoms and improves qual

ity of life in patients With active Crohns disease. TWo-thirds of enrolled patients achieved remission at some point during LDN treatment. Another ?nding in this trial Was the fairly rapid onset of effect from LDN in that by four Weeks there Was signi?cant improvement. Corticosteroids may be effec
50

day. Another advantage of LDN is the once-a-day dosing Which may improve patient compliance.
Ulcerative Colitis

In the folloWing detailed example embodiment, the opioid

antagonist naltrexone is shoWn to reduce in?ammation of the
55

tive in decreasing symptoms of Crohns patients in 7-10 days,

but other medications such as the immunomodulators (e.g.,

boWel in a chemically-induced mouse model of ulcerative

colitis. The dextran sulfate sodium (DSS) model of experi

mental colitis in mice is a common pre-clinical model for IBD due to its ease of administration and e?icient and reversible

aZathioprine and 6-mercaptopurine) may take 3-4 months to

demonstrate improvement in symptoms. Often symptoms

recur Within one month after discontinuing corticosteroids or

induction of symptoms. The addition of DSS to drinking

60

aminosalicylates. HoWever, in the present study continued

improvement in CDAI scores and quality of life Was reported even four Weeks after discontinuing LDN.

Water induces hematocheZia, Weight loss, intestinal shorten

ing and in?ltration of neutrophils, thus serving as a model for

human in?ammatory boWel diseases. BreakdoWn of epithe

lial barrier function in DSS-treated mice leads to an induction
65

Another ?nding in this pilot study Was that LDN improved

the quality of life of subjects With active Crohn s disease. The baseline value on the IBDQ Was similar to that reported in other clinical trials indicating that our subject group did not differ from those used in other studies. Statistical analysis

of pro-in?ammatory cytokines, Which are thought to play a central role in disease progression. Cytokines can both sup press apoptosis and drive activation of immune cells that
contributes to a chronic state of in?ammation. Treatments

US 7,879,870 B2
13
aimed at reducing this excessive in?ammatory response have

14
minimiZe the impact of variation among animals, RNA from
individual mice Was pooled Within each treatment group.

demonstrated therapeutic promise in DSS models. Therefore,

DSS-treated mice are particularly suitable for proof-of-con cept studies of novel IBD therapeutics and treatments that may reduce the cytokine-induced in?ammatory state. The effect of naltrexone on the progression of DSS-induced colitis in mice Was examined. To more accurately model therapeutic invention in established colitis, chemical induction preceded
the initiation of treatment. The ability of naltrexone to reverse the acute in?ammatory responses induced by DSS Was ana

Arrays Were run in duplicate and three pairWise comparisons Were performed to identify genes for Which expression Was

signi?cantly altered (p<0.05) in colitis and by subsequent

naltrexone treatment. RNA from Normal+saline, DSS+6.3 ug/kg NTX, and DSS+350 ug/kg NTX Were each tested ver sus DSS+saline. Data Were analyZed With GenSpring 6 soft

lyZed at the level of Whole animal symptoms, tissue histology, and RNA pro?ling. Six to eight-Week-old male C57 black/6] mice (Charles River Laboratories, Inc., Wilmington, Mass.) Were randomly
allocated into one of tWo groups of 24 mice each. Food and

Ware (Agilent Technologies, Palo Alto, Calif.). RNA (18S and 28S bands) Was visualiZed using the Agilent

2100 BioanalyZer (Agilent Technologies, Santa Clara, Calif.)

and concentrations Were adjusted. First strand cDNA Was

then produced from 1.0 pg of total RNA using random hex amer primers and the SuperScript III Reverse Transcription

Water Were provided ad libitum. The ?rst group (Normal) received normal drinking Water and the second group (DSS) received Water containing 2% dextran sulfate sodium having

kit (Invitrogen). The concentration and quality of resulting cDNA Was quanti?ed and analyZed using the Agilent 2100 BioanalyZer or spectroscopically With the NanoDrop

a molecular Weight of 40,000 (TdB Consultancy AB, Upp sala, SWeden) for six days folloWed by normal Water for three
additional days and necropsy on day 9 (FIG. 5A). Each mouse
Was housed in an individual cage for accurate measurement of

ND-1000 (NanoDrop Technologies, Wilmington, Del.).

20

Samples Were standardized to 30 ng/uL and 60 ng of cDNA per sample Was then used as a template for real-time PCR

food and Water intake. Animal Weight, Water intake, and food consumption Were measured daily. Bedding Was changed and fresh stool pellets Were collected daily and analyZed for occult blood. The colitis disease activity index (DAI) Was calculated for each mouse according to the system estab

using a SYBR Green Master Mix (Qiagen, Valencia, Calif.). 18S rRNA primers (Eurogentec, San Diego, Calif.) and the

folloWing gene-speci?c primer sequences obtained from

PrimerBank (pga.mgh.harvard.edu/primerbank) Were used:
25

lished by Murthy and colleagues. See, Murthy, et al., Treat

ment of dextran sulfate sodium-induced murine colitis by

beta-actin, 6671509a; IL-5, 6754336a; IL-6, 13642311a; IL-12, 6680395a; STAT3, 13277852a; STAT4, 6755670a; Muc2, 3452503a2; TFF3, 6755773a1; Palladin, 9828173a1;
and TGF-beta BP, 7305243a1. To exclude the possibility of genomic DNA contamination, control reactions With no cDNA template Were also performed for each gene-speci?c primer set. PCR ampli?cation and analysis Were performed

intracolonic cyclosporin, Dig. Dis. Sci. 38, 1722-34 (1993).

Overt changes in stool consistency Were rarely discerned so a modi?ed DAI Was calculated based on percent Weight loss and stool hemoccult or presence of gross bleeding. Mice in each group (Normal or DSS) Were randomly sub divided into three treatment groups of eight mice each. After 72 hours, mice Were treated once daily for six consecutive

30

With the Applied Biosystems Sequence Detection System 7300 using the Relative Quanti?cation (ddCt). Ampli?cation
data for the genes of interest Were normaliZed to 18S Within
35

days With a subcutaneous injection (0.1 mL) of one of the

each individual reaction. Triplicate reactions Were performed and the resultant data from multiple runs Were averaged.
Results Were calculated as meaniSEM. Statistical com

folloWing: saline (control), 6.3 ug/kg naltrexone (NTX), or 350 ug/kg NTX (Sigma Chemicals, St. Louis, Mo.). On day 9,
all animals Were necropsied and their colons resected. At necropsy, the entire colon Was excised, measured in
40

parisons Were performed betWeen NTX treatment sub -groups and their corresponding saline control, as Well as betWeen the Normal and DSS groups, comparing corresponding treat

length, and bisected into proximal and distal portions. The

proximal and distal colons Were additionally divided for RNA extraction and histology. Each histology specimen Was ?xed in 10% neutral buffered formalin, paraf?n embedded and
45

ments. Parametric analyses Were performed (Minitab 13, State College, Pa.) using a modi?ed Bonferroni method to correct for multiple comparisons to controls. Over the nine day course of study, animals given untreated

sectioned for hematoxylin and eosin (H&E) staining.

Specimens Were examined microscopically and scored based upon the criteria established by Williams et al. by an investi gator blinded to the treatment groups. Williams, et al., Enhanced survival and mucosal repair after dextran sodium sulfate-induced colitis in transgenic mice that overexpress

drinking Water exhibited steady Weight gain (FIG. 5A, upper

panel) While DSS mice shoWed Weight loss beginning betWeen days 4 and 6 (FIG. 5A, loWer panel). Steady Weight
loss continued in the DSS mice until necropsy (day 9). With
50

naltrexone treatment, the Weight loss tended to be less com

groWth hormone, Gaslroenlerology 120, 925-37 (2001).

Brie?y, a representative longitudinal section from each
mouse Was scored at six random ?elds for in?ammation

pared to DSS+saline; hoWever, the DSS+6.3 ug/kg NTX mice only reached statistical signi?cance on day 6 (p:0.02). Among the three DSS sub-groups, no signi?cant difference in
Weight Was evident at necropsy, When the animals had been
55

severity, extent of in?ammation (mucosa, submucosa, trans mural) and crypt damage. Each of these scores Was Weighted
to re?ect the percent involvement of the overall section and the Weighted scores from each of the six ?elds Were averaged
to achieve an overall in?ammation score for each mouse.

off DSS for three days.

Naltrexone treatment alone had no effect on food con

sumption (FIG. 5B). HoWever, DSS mice, With or Without

60

Total RNA Was extracted from the distal colon samples

NTX treatment, had signi?cantly decreased total food con sumption (FIGS. 5B and 5C). Total Water intake Was also reduced in DSS mice compared to mice receiving normal
Water. Among the Normal mice, a reduction in Water con

(using TRIZOL available from Invitrogen Corp., Carlsbad,

Calif.; TRIZOL is a trademark of Molecular Research Cen

ter, Inc. of Cincinnati, Ohio) and subjected to microarray analysis using a 10K mouse microarray (MWG Biotech, High
Point, NC.) and a Packard Biosciences ScanArray 4000KL machine (Functional Genomics Core Facility of the Section of Research Resources, Penn State College of Medicine). To
65

sumption Was noted in those treated With 6.3 ug/kg NTX relative to saline (FIG. 5C). In FIGS. 5B and 5C, asterisks

indicate signi?cantly different values betWeen corresponding DSS and Normal (no DSS) treatment groups (*p<0.025; **p<0.005; n:7-8). Signi?cant differences betWeen saline
and NTX-treated mice are indicated by a T(p<0.025).

US 7,879,870 B2
15
To monitor disease progression, a disease activity index (DAI) Was assessed daily for each mouse. Modi?ed DAI scores for all Normal mice (both saline and NTX-treated) shoWed no evidence of colitis (FIG. 6, upper panel). As With

16
be up-regulated in DSS-induced colitis, Were increased in DSS+saline animals in comparison to Normal controls (FIGS. 8B and 8C). The increase in IL-6 mRNA Was 73-fold,
While the signi?cant increase in IL-l2 Was more modest

Weight loss, the DSS mice developed colitis symptoms, including hemoccult-positive stools and increased DAI scores, by day 4 Which continued to increase through day 8
(FIG. 6, loWer panel). A reduction in these DAI scores Was

(three-fold). Upon treatment With naltrexone, levels of IL-6

and IL-l2 Were greatly reduced in DSS mice. For IL-l2,
naltrexone treatment restored mRNA expression to that seen in the colitis-free, Normal mice. The reduction in IL-6 Was

evident With naltrexone treatment. On day 6, DSS+350 ug/kg NTX animals had signi?cantly loWer (55%) DAI scores than DSS+saline mice (p:0.0l5). In FIG. 6, a signi?cant differ ence betWeen DSS+350 ug/kg NTX-treated mice and the

also signi?cant, although levels Were not completely restored

to those seen in the colitis-free animals.

To assess Whether the alteration in the expression of pro

corresponding DSS+saline group is indicated (l"p<0.05; n:7

in?ammatory cytokines IL-6 and IL-l2 mRNAs had func tional consequences, the mRNA for doWnstream signaling
intermediates Was also measured by real-time RT-PCR. Both

8).
Reduced colon length, another indicator of DSS-induced colitis, Was also evident in all DSS animals (Normal+saline: 9.321039 cm, meanzSEM; DSS+saline: 6.95:0.43;
p:0.002). DSS-mice treated With naltrexone had more nor

STAT3, doWnstream of IL-6, and STAT4, doWnstream of

IL-l2, Were increased in DSS+saline animals (2.20-fold and

8.03-fold, respectively), again indicating a pro-in?ammatory

response had been induced (FIGS. 8B and 8C). HoWever,
20

mal colon lengths, although they Were still shorter than in the untreated animals.
Histological in?ammation scores of the distal colon con

both STAT3 and STAT4 mRNA levels Were decreased by greater than 50% after naltrexone treatment. In fact, levels of STAT3 mRNA in the naltrexone-treated animals Were statis

?rm that DSS induced an in?ammatory state (FIG. 7A). No differences in colonic in?ammation in Normal+saline and Normal+NTX animals Were evident histologically. This indi cates that naltrexone alone did not alter the mucosal integrity of the colon. All DSS animals had increased in?ammation scores, and exhibited crypt damage and increased leukocyte in?ltration (FIG. 7B). HoWever, the DSS+NTX animals had a dose-dependent decrease in in?ammation and damage, as evidenced by loWer in?ammation scores in comparison to

tically indistinguishable from colitis-free, Normal controls.

Although STAT4 mRNA levels Were signi?cantly reduced by naltrexone treatment, they Were still slightly higher than in
25

the Normal mice.

In FIG. 8, histogram columns represent the relative quan

tity (RQI2-DDCT) for each calibrator/target pair. Bars rep

resent a 95% con?dence interval (CI) around the relative
30

animals given DSS+saline (FIG. 7A). In FIG. 7A, asterisks indicate signi?cantly different values between corresponding DSS and Normal (No DSS) treatment groups (*p<0.025;
**p<0.005; n:7-8). While this trend did not reach statistical signi?cance, the DSS+350 ug/kg NTX mice had improved crypt architecture and feWer invading leukocytes than Were observed in the DSS+saline mice (FIG. 7B). In FIG. 7B,
representative H&E sections of distal colon are shoWn. Com
35

quantity, and signi?cance is based on non-overlapping CIs. Relative RNA levels for (A) beta-actin and the cytokine IL-5; (B) cytokine IL-l2 and its doWnstream effector STAT4; and

(C) cytokine IL-6 and its downstream effector STAT3, reveal signi?cant DSS-induced elevations in pro-in?ammatory mediators, and signi?cant restoration toWard normal levels
When animals Were treated With naltrexone.

pared to the normal appearance of healthy murine colon (no DSS+saline; top), leukocyte in?ltration and an absence of
normal crypt architecture are evident in the DSS+saline mice

40

(middle). Improved architecture and less in?ammation are clearly discemable in DSS mice treated With 350 ug/kg nal trexone (bottom of FIG. 7B). Differences in distal colon gene expression betWeen Nor mal+saline, DSS+saline, and DSS+NTX mice Were initially assessed using a murine spotted oligonucleotide array. Out of
9800 genes on the arrays, 506 genes Were initially identi?ed

This study is the ?rst to report improvement of colitis in a murine model upon treatment With an opioid antagonist. Nal trexone treatment resulted in a rapid mitigation of colitis symptoms in DSS mice, While in the absence of colitis, it had no signi?cant impact. DSS Was administered for three days prior to either saline or naltrexone injections, emulating a condition of established boWel in?ammation preceding treat ment. Animals With DSS in their drinking Water exhibited

signi?cant Weight loss and symptoms of colitis; hoWever,

45

Within three days of treatment With 350 ug/kg naltrexone,

both Weight loss and disease symptoms Were decreased. In?ammation Was also improved as indicated by histology at the studys conclusion. Indeed, the differences in the histo logical in?ammation scores may have been more pronounced if evaluated at an earlier time point While DSS Was still being administered. Cessation of DSS on day 6 may have conceiv

as being signi?cantly changed (p<0.05) in DSS+saline mice When compared to Normal+saline mice (data not shoWn).
Among the most differentially expressed genes Were mucin

50

(Muc2), trefoil factor (TFF3), and TGF-beta binding protein.

Naltrexone treatment (either 6.3 or 350 ug/kg) restored the levels of these mRNAs in DSS animals to varying extents.

ably alloWed for partial recovery, mollifying potentially

greater signi?cance of the naltrexone treatment. A robust, pro-in?ammatory response characteristic of IBD
55

HoWever, upon subsequent validation by quantitative RT

PCR, the levels of these three mRNAs in DSS+saline and DSS+NTX animals Were not signi?cantly different from Nor mal animals.

Was further con?rmed at the molecular level. Signi?cant

elevations in the gene expression of pro-in?ammatory cytok

ines IL-6 and IL-l2 Were observed With DSS. Molecular

Because naltrexone reduced the in?ammatory histology of DSS-induced colitis, the expression of several genes of inter est, including both cytokines and doWnstream mediators, Was examined by real-time RT-PCR. Expression of beta-actin, cytokines IL-5, IL-6, IL-l2 and transcription factors STAT3
and STAT4, Were assessedusing 18S rRNA as an endogenous control. The expression of beta-actin and IL-5 Were not sig

evidence also indicated that subsequent naltrexone treatment had a dramatic impact on these pro-in?ammatory signaling
60

pathWays, signi?cantly decreasing expression of IL-6 and

IL-l2 in DSS mice to normal or near-normal levels. These

65

decreases in cytokine expression also proved to be func tionalireducing the expression of the doWnstream signaling molecules, STAT3 and STAT4. By moderating the over stimulation of immune responses and restoring cytokine lev
els, naltrexone treatment alloWed a more normal mucosal structure to reappear. By this means, naltrexone treatment

ni?cantly changed by either DSS+saline or by DSS+NTX (FIG. 8A). By contrast, cytokines IL-6 and IL-l2, knoWn to

US 7,879,870 B2
17
reversed the increase in both the molecular markers of disease

18
6. The method according to claim 1, Wherein said boWel
disease is Crohns disease or ulcerative colitis.

and the physiological symptoms of colitis induced by DSS.

Since current therapies, such as anti-TNF-ot monoclonal

antibodies, are designed to eliminate pro-in?ammatory cytokines, these therapies carry an increased risk of infection due to immune suppression. HumaniZed monoclonal anti
bodies also shoW diminished e?icacy over time and have

signi?cant secondary complications, decreasing their suit

ability for long-term use. Because naltrexone doWn-regulates but does not eliminate pro-in?ammatory cytokines, naltrex
one therapy may have feWer undesirable side-effects than

7. The method according to claim 1, Wherein said pharma ceutically acceptable salt is a hydrochloride salt thereof. 8. The method according to claim 1, Wherein said opioid antagonist is provided as a pharmaceutical composition. 9. The method according to claim 8, Wherein said pharma ceutical composition is administered once per day in the
evening or at bedtime.

10. The method according to claim 8, Wherein said phar maceutical composition is administered once per day in the

currently used agents. Additionally, the versatility of naltrex

one for oral administration presents advantages in patient

morning or after Waking from sleep. 11. The method according to claim 8, Wherein said phar
maceutical composition is formulated as a solid dosage form suitable for oral administration. 12. The method according to claim 11, Wherein said solid dosage form is an immediate release formulation comprising an opioid antagonist and an excipient.
20

compliance.
While the invention has been described With reference to

preferred embodiments, it Will be understood by those skilled

in the art that various changes may be made and equivalents may be substituted for elements thereof Without departing from the scope of the invention. In addition, many modi?ca
tions may be made to adapt a particular situation or material

to the teachings of the invention Without departing from the essential scope thereof. Therefore, it is intended that the invention not be limited to any particular embodiments dis closed as the best mode contemplated for carrying out this invention, but that the invention Will include all embodiments falling Within the scope of the appended claims. The invention claimed is: 1. A method for treating a boWel disease comprising administering to a subject in need thereof a therapeutically effective dose less than 50 mg of an opioid antagonist effec
tive to treat a boWel disease in said subject, Wherein said boWel disease is characterized by in?ammation or ulceration

25

13. The method according to claim 12, Wherein said excipi ent is selected from the group consisting of sucrose, cellulose, and combinations thereof. 14. The method according to claim 8, Wherein said phar maceutical composition is formulated as a topical dosage form suitable for topical administration. 15. The method according to claim 8, Wherein said phar maceutical composition of a therapeutically effective dose less than 50 mg is formulated as a liquid dosage form suitable for oral administration. 16. The method according to claim 8, Wherein said liquid

30

dosage form comprises an opioid antagonist and a liquid

carrier.

of the intestinal Wall and Wherein the opioid antagonist is selected from the group consisting of naltrexone, nalmefene,

naloxone and pharmaceutically acceptable salts thereof, and

combinations thereof. 2. The method according to claim 1, Wherein said boWel disease is further characteriZed by abdominal pain or discom fort, abnormal boWel movement frequency, intestinal steno sis or ?stuliZation, perianal discomfort or pruritis, or abnormal

35

40

stool consistency.
3. The method according to claim 1, Wherein said boWel disease is further characteriZed by in?ammation or ulceration
of the small intestine or colon.

17. The method according to claim 8, Wherein said liquid carrier comprises Water. 18. The method according to claim 1, Wherein said thera peutically effective dose less than 50 mg is from about 1.75 mg to about 4.5 mg of said opioid antagonist. 19. The method according to claim 1, Wherein said thera peutically effective dose less than 50 mg is from about 1.75 mg to about 3 mg of said opioid antagonist. 20. A method of pharmaceutical treatment comprising orally administering to a human subject having Crohns dis
ease or ulcerative colitis a therapeutic pharmaceutical com

position once per day in the evening or at bedtime, Wherein

said pharmaceutical composition comprises from about 3 mg

45

4. The method according to claim 1, Wherein said boWel disease is further characteriZed by the presence of a ?stula. 5. The method according to claim 1, Wherein said boWel

to about 4.5 mg of naltrexone, nalmefene, naloxone, or a hydrochloride salt thereof in an immediate release solid dos age formulation.

disease is in?ammatory boWel disease (IBD).

US 20070259939Al

(19) United States (12) Patent Application Publication (10) Pub. No.: US 2007/0259939 A1 Stebbing (43) Pub. Date: NOV. 8, 2007
(54)
USING NALTREXONE AS A MULTI-PURPOSE HEALTH SUPPLEMENT TO IMPROVE THE HUMAN CONDITION AND PREVENTING MULTIPLE DISEASES AND INFIRMITIES BY STIMULATING IMMUNE SYSTEM VITALITY AND ROBUSTNESS Publication Classi?cation

(51)

Int. Cl. A61K 31/485

(2006.01)

(52)

us. c1. ..................................................... .. 514/410

(75) Inventor:

Franklin Leroy Stebbing,

(57)

ABSTRACT

Norfolk, NE (U S)
Correspondence Address:
BANNER & WITCOFF, LTD. TEN SOUTH WACKER DRIVE, SUITE 3000 CHICAGO, IL 60606

Naltrexone has been used to ?ght existing infections, ai?ic

tions, and substance abuse. Embodiments of the current invention provide naltrexone as a preventative medicine

taken daily, usually in the late evening for greater efficacy.

It is taken in a loW dose like a vitamin supplement and may

prevent many diseases by stimulating the immune system.

The increased endorphin levels resulting from the use of loW dose naltrexone may also reduce the need for pain relief

(73) Assignee:

ACCELERATED TECHNOLOGIES, Norfolk, NE

medications, plus counter tendencies toWard experiencing

undesirable moods, such as depression or anger. Increased

(Us) (21) Appl. No.: (22)

Filed:

11/550,742
Oct. 18, 2006

feelings of Well-being produced by naltrexone may also have far-reaching eifects, as suggested by studies shoWing
that people With positive attitudes recover more quickly from serious illnesses, for example. In another aspect of the invention, naltrexone may be used to prevent substance addiction before it ever starts, by blocking the internal positive response that normally results from the use of chemical substances.

Related US. Application Data

(60) Provisional application No. 60/797,587, ?led on May

4, 2006.

US 2007/0259939 A1

Nov. 8, 2007

USING NALTREXONE AS A MULTI-PURPOSE HEALTH SUPPLEMENT TO IMPROVE THE HUMAN CONDITION AND PREVENTING MULTIPLE DISEASES AND INFIRMITIES BY STIMULATING IMMUNE SYSTEM VITALITY AND ROBUSTNESS CROSS REFERENCE TO RELATED APPLICATIONS

blocking the receptors that provide the stimulatory effect felt

from alcohol. As a result of the lack of positive reinforce ment, there is no internal incentive to drink alcohol. Naltr exone is not knoWn to have any important side effects at loW

[0001] This application claims priority to US. Provisional Application No. 60/797,587 ?led May 4, 2006.
FIELD OF THE INVENTION

dose levels. The FDA approved dosage siZe for treatment of alcohol dependency is a 50 mg tablet, once daily. While taking naltrexone at this dosage level, it is suggested that a counselor should be involved for support and advice and supervision. It also can be given in a sloW release form by injection or surgical implant if the patient is incapable or cannot be depended upon to take the medicine regularly.
SUMMARY OF THE INVENTION

[0006]

Previously, naltrexone had only been used to ?ght

[0002]

The invention relates to health supplements, in

existing infections or af?ictions, or to prevent recurrence. In

particular supplements for the prevention of diseases and

in?rmities and for the reduction of susceptibility toWards chemical addiction.
BACKGROUND OF THE RELEVANT ART

contrast, embodiments of this invention provide naltrexone as a preventative medicine taken daily, usually in the late evening for greater ef?cacy. It is taken like a vitamin supplement and may prevent many of the diseases that tend
to be susceptible to a strong immune system.

[0007]
[0003] Traditional medicine has long been focused on the cure of a?lictions after they are recognized by the onset of symptoms. Aspects of this invention are dedicated to the proposition that it is better to prevent diseases then to cure
them after they have become established. There are numer ous health supplements and vitamins available noW that are

The increased endorphin levels resulting from the

use of loW dose naltrexone may also reduce the need for pain
relief medications as Well as counter tendencies toWard

experiencing undesirable moods, such as excessive depres sion or anger. Increased feelings of Well-being may also

have far-reaching effects, as suggested by studies shoWing

that people With positive attitudes recover more quickly from serious illnesses, for example. In addition, any pre vention of sickness Would have the additional bene?ts of decreasing the use of medications With unWanted side effects
and loWering health care costs.

just for the maintenance and/ or enhancement of good health and vitality. More and more people are desirous of holding onto vigor and vitality as they age, such as baby boomers. Many people are determined to maintain their health and vitality for as long as it is possible to do so, and in?rmity is
often not accepted as inevitable. [0004] Naltrexone has been used in loW dose form since at least the mid 1980s to cure or stabiliZe a groWing list of ailments. It is used for the treatment of a number of serious diseases, such as some cancers, HIV, AIDS and multiple sclerosis. Studies have shoWn naltrexone to have a positive effect in as many as 65% of cancer patients, for example

[0008] In aspects of the invention, naltrexone may be used to prevent substance addiction before it ever starts, by taking it daily. This could result in great personal and ?nancial savings by reducing the occurrence of addiction and thus the need for rehabilitation programs. Unlike existing medica
tions such as Antabuse, a drug that Will induce nausea or

even unconsciousness if it is taken folloWed by alcohol consumption, naltrexone is more effective When taken prior
to drinking alcohol because it reduces or removes the desire to drink.

stopping the spread of tumors, as Well as causing remission in 30% of the patients. LoW dose naltrexone functions by blocking opioid, or endorphin, receptors in various types of

cells. By blocking opioid receptors, naltrexone also blocks

the reception of the opioid hormones that our pituitary and

adrenal glands produce: beta-endorphin and metenkephalin.

Many body tissues have receptors for these endorphins and enkephalins, including virtually every cell of the bodys
immune system. When the receptors are blocked, the body signals an increase in endorphin production and the level of endorphins rises in reaction. An increase in endorphins
causes the immune system to become stimulated and there fore more effective at ?ghting infection. The increased

DETAILED DESCRIPTION OF EMBODIMENTS OF THE INVENTION

[0009]

One embodiment of the invention is a method of

using loW doses of naltrexone to block endorphin receptors

in various body cells of a human or animal patient. When the receptors are blocked, the body signals an increase in

endorphin production and the level of endorphins rises in

reaction. The method may comprise a daily dose of naltr exone administered to the patient, for example betWeen

endorphin level may also dull or kill pain, and often results in a feeling of Well-being because endorphins are natural mood elevating chemicals in the brain. In contrast, stress
may cause a marked reduction in endorphins, Which often

approximately 0.5 and 5 milligrams of naltrexone, and preferably betWeen 1.75 and 4.5 milligrams of naltrexone. An aspect of the invention provides administration of the daily dose of naltrexone in the late evening hours, such as
betWeen 6:00 pm and 2:00 am, so as to block the opioid

does not correct itself naturally, thus resulting in loWered immune defenses susceptible to a variety of illnesses, such
as autoimmune diseases or cancer. Cancer has occasionally

receptors at a time When endorphins are naturally produced

by the body s circadian rhythm, usually in the early morning

betWeen 2:00 am and 4:00 am, regardless of Wakefulness or

been observed in patients after about tWo to six years folloWing a period of extreme stress. [0005] With respect to the treatment of alcohol addiction, naltrexone may be effective because it eliminates the neural

sleep. The brief blockade of opioid receptors betWeen 2 am.

and 4 am. may produce a prolonged stimulation of the

positive reinforcement normally experienced in the brain by

immune system by causing an increase in endorphin and enkephalin production. In fact, tests have shoWn that taking

US 2007/0259939 A1

Nov. 8, 2007

small doses of naltrexone each night has resulted in much

feeling of Well-being, naltrexone may reduce anti-social

behavior With more incidences of positive social interac tions. [0013] Naltrexone is already knoWn to help cure addictive

higher levels of beta-endorphins circulating in the blood in

the following days, providing as much as a 200-300%

increase in patients Who initially had de?cient endorphin

levels. The naltrexone may be taken as a tablet, capsule or

diluted in liquid, as long as it is present in a readily bioavailable form. [0010] An embodiment of the invention is a method to stimulate the immune system of a patient prior to the onset of an illness via a daily dose of naltrexone that is generally in the range of 0.5 to 5 milligrams per dose, and again

behaviors, such as alcoholism, opiate, narcotic and tobacco addiction. With respect to the treatment of alcohol addiction,
naltrexone may be effective because it eliminates the neural

positive reinforcement normally experienced in the brain by blocking the receptors that provide the stimulatory effect felt
from alcohol. As a result of the lack of positive reinforce
ment, there is no internal incentive to drink alcohol. In a

preferably betWeen about 1.75 and 4.5 milligrams per dose.

The resulting increase in endorphins thus causes a stimula

similar manner, in another embodiment of the invention, the

daily administration of loW dose naltrexone may greatly

reduce or eliminate the tendency to engage in addictive

tion of immune system function With marked improvement in disease ?ghting capability. In an aspect of the invention, the administration of a daily dose of naltrexone, for example
in the late evening, may prevent the onset of an illness in a

behavior by preventing its onset. This may be accomplished

by diminishing any potential internal reWards for using

chemical substances in the ?rst place. The loW dosage of naltrexone, for example around 4.5 mg, has been found to greatly inhibit alcohol cravings. The diminished desire for alcohol resulting from a loW dose of naltrexone may be effective for more than 24 hours, thus for example a dose
administered at 7:00 pm one evening may continue to inhibit

human or animal patient. The method includes taking loW

doses of naltrexone prior to potentially being exposed to

infectious conditions, With or Without additional medica tions or vaccinations to cause antibody production. An

increase in endorphins resulting from the blockage of endor phin receptors by the naltrexone causes the immune system
to become more effective at ?ghting infection during its early stages. The use of naltrexone as taught here may

alcohol cravings throughout the folloWing evening. Aperson

in need of curbing an alcoholic appetite may take a pill, either before or along With the drink, and discover no desire
for a second or third drink. To further eliminate an alcoholic

improve immune system function, resulting in a greatly

enhanced ability to fend off debilitating diseases such as

appetite, a series of 1.5 mg naltrexone doses may be taken

many forms of cancer, AIDS, HIV, multiple sclerosis, ALS, AlZheimers Disease, chronic fatigue syndrome, emphy sema, Parkinsons Disease, primary lateral sclerosis, psoria sis, rheumatoid arthritis, sarcoidosis and systemic lupus. For
example, cancer cells are routinely produced in the body, but a healthy immune system generally kills them as they occur. In fact, endorphins added to groWing human cancerous
tissue in a laboratory Petri dish have been observed to kill
the cancerous tissue. In contrast to Waiting for a disease to

Without alcohol, starting for example at about 6:00 pm. Three doses, for a total of 4.5 mg naltrexone, may be taken,
for example tWo or three doses together or single doses over a period of time, such as one per hour, as provides the most effective result for the individual. [0014] It is knoWn that it is very important to have a

positive attitude and outlook When battling and treating

serious illnesses, such as cancer. The outcome is much better

if the patient is upbeat and optimistic. This is similar to What

happens to those Who take loW doses of naltrexone to boost endorphin levels to combat diseases. The loW doses of

become established and then taking steps to cure it With

medications like naltrexone and/or other means such as

naltrexone raise the level of endorphins, Which eventually

leads to an increase in immune system function. Either Way,

surgery or chemotherapy, the disease may be prevented. Additionally, if an illness does become established, the daily naltrexone therapy may reduce or eliminate the need for
various medications to overcome the illness because of the

naturally or naltrexone induced, the increased levels of endorphins result in a better mood and an improved, enhanced and more robust immune system function, Which equates to an improved prognosis due to a greater ability of
the immune system to overcome and defeat a disease. If a

enhanced natural ability of the immune system.

[0011] An additional aspect of the invention is a method to alleviate or reduce pain in a human or animal patient comprising a daily administration of loW dose naltrexone. A naltrexone dose of betWeen about 0.5 and 5 milligrams may

patient does not have a naturally optimistic attitude, naltr

exone may help provide a good disposition With the same

bene?cial results, by arti?cially stimulating the immune system.

[0015] Variations and modi?cations of the foregoing are Within the scope of the present invention. It should be understood that the invention disclosed and de?ned herein
extends to all alternative combinations of tWo or more of the individual features mentioned or evident from the text. All

be used, for example taken during the late evening such as

from 6:00 pm to 2:00 am, as described above. The increased

endorphin level may dull or kill pain, as endorphin levels naturally increase When injury occurs. Because naltrexone stimulates an endorphin increase, naltrexone therapy may reduce aches and pains. [0012] The employment of a daily loW dose of naltrexone
may prevent one or more psychological problems in a

of these different combinations constitute various alternative aspects of the present invention. The embodiments

described herein explain the best modes knoWn for practic

ing the invention and Will enable others skilled in the art to
utiliZe the invention. The claims are to be construed to

human, in an embodiment of the invention. This is due to the

resulting increase of endorphins, Which are natural mood elevating chemicals in the brain. An increased level of endorphins may preclude or prevent the onset of some mental depression or anger, for example, because endor phins are natural anti-depressives and mood enhancers. As a result of the absence of depression and improved mood and

include alternative embodiments to the extent permitted by the prior art. For example, endorphins may be administered
directly Without the use of naltrexone such as in those cases

Where a patient is not able to produce endorphins by

naltrexone stimulation, or Where the use of naltrexone is

US 2007/0259939 A1

Nov. 8, 2007

prevented by other factors. For instance, because naltrexone

is an opiate agonist, it may not be appropriate to administer

naltrexone to a patient using opiates for pain medication.

[0016] Various features of the invention are set forth in the

following claims.
What is claimed is:
1. A method to increase endorphin levels in a human or

animal patient comprising a daily administration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 2. The method according to claim 1 Wherein the naltrex one is administered to the patient during an evening hour. 3. The method according to claim 2 Wherein the evening hour is betWeen approximately 6:00 pm and approximately
2:00 am.

11. The method according to claim 9 Wherein the chemi cal substance is an opiate. 12. The method according to claim 9 Wherein the chemi cal substance is alcohol. 13. The method according to claim 9 Wherein the chemi cal substance is nicotine. 14. A method to reduce physical pain in a human or animal patient comprising a daily administration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 15. The method of claim 14 Wherein the naltrexone is

administered to the patient during an evening hour. 16. The method of claim 15 Wherein the evening hour is betWeen approximately 6:00 pm and approximately 2:00
am.

4. A method to stimulate the immune system of a human or animal patient prior to the onset of an illness comprising a daily administration of a dose of naltrexone. 5. The method according to claim 4 Wherein the dose of naltrexone is betWeen about 0.5 mg and about 5 mg. 6. The method according to claim 4 Wherein the naltrex one is administered to the patient during an evening hour. 7. The method according to claim 6 Wherein the evening

17. A method to prevent the onset of illness in a human or

animal patient comprising a daily administration during an

evening hour of a dose of betWeen about 0.5 mg and about 5 mg naltrexone. 18. The method of claim 17 Wherein the evening hour is

betWeen approximately 6:00 pm and approximately 2:00

am.

hour is betWeen approximately 6:00 pm and approximately

2:00 am.

19. A method to prevent one or more psychological

8. Amethod to prevent addiction of a human or an animal

problems in a human patient comprising a daily adminis

tration of a dose of betWeen about 0.5 mg and about 5 mg naltrexone.
20. The method of claim 19 Wherein the one or more

patient to a chemical substance comprising a daily admin

istration of a dose of naltrexone.

9. The method according to claim 8 Wherein the naltrex

one is administered prior to any use of the chemical sub
stance.

psychological problems are selected from the group com

prising depression, anger and anti-social behavior.

10. The method according to claim 8 Wherein the dose of naltrexone is betWeen about 0.5 mg and about 5 mg.