EUROPEAN JOURNAL OF DRUG MIITABOLISM AND PHARMACOKINETICS, 1990, Vol. 15, No.2, :pp.

83-93

Transport of peptide and protein drugs across biological

membranes

J.e. VERHOEF, H.E. BODDE, A.G. de BOER, J.A. BOUWSTRA,

H.E. JUNGINGER, F.W.H.M. MERKUS and D.D. BREIMER
Center for Bio-Phannaceutical Sciences, Leiden University,
Leiden; The Netherlands

Keywords: Peptides, protein drugs, biological membranes.

SUMMARY
The transport characteristics of peptide and proteins drugs across various epithelial membrane barriers are outlines. These include transport
through the intestinal, buccal, nasal and pulmonary absorptive mucosae, as well as transdermal penetration. Because peptides and proteins are
hydrophilic and high molecular weight compounds, they commonly show minor permeability across the mentioned biological membranes. In order
to improve their transport properties and thereby their systemic bioavailability, several strategies can be undertaken, such as the synthesis of stabil-
ized and lipophilic analoques, the application of absortion enhancers and protease inhibitors, and the design of suitable dosage forms (e.g., Iiposomes,
biodegradable nanocapsules, bioadhesive microspheres).

INTRODUCTION ministered peptides and proteins, as well as of possible

Peptides and proteins are endogenous compounds
that regulate endocrine and other physiological
processes in the body. Because of their high potency
and suspected low toxicity, the interest of the phar-
maceutical industry as well as academia is now grow-
ing massivelyto develop peptides, their derivatives and
proteins as drugs for diagnostic and therapeutic pur-
poses. This is also encouraged by the present pos-
sibilities to synthesize oligopeptides on a large,
preparative scale, and to produce polypeptides and
proteins (e.g., insulin, growth hormone, and inter-
ferons) by recombinant DNA technology. Nonethe-
less, serious handicaps still exist with respect to the
delivery and disposition of peptide and protein drugs:
these substances generally have to be administered by
injection since they show minimal bioavailability after
oral administration, due to their instability in the
gastrointestinal tract, poor absorption characteristics,
and.their rapid and extensive biotransformation in the
body. The present paper will focus on the transport of
peptide and protein drugs across various epithelial
membane barriers following administration by various
routes. An overviewwill be given of the metabolic fate
and intestinal transport mechanisms of perorally ad-
Send reprintrequeststo : Dr. J.e.Verhoef, Center for Bio-
Pharmaceutical Sciences, Leiden Univercity, P.O. Box
9502, 2300 RA Leiden, The Nederlands
strategies to improve the peroral bioavailability of
these compounds. Moreover, recent promising results
regarding the transport of peptides and proteins across
alternative absorptive mucosae will be discussed, the
emphasis being on the transport through buccal, rec-
tal, nasal and pulmonary epithelial mucosae, and on
percutaneous transport. Progress in this challenging
field of research will also be illustrated with results ob-
tained in our Center for Bio-Pharmaceutical Sciences
at Leiden University.
GASTROINTESTINAL TRANSPORT
The problems which make the oral route incon-
venient for systemic delivery of peptide and protein
drugs are the degradation by the strong acidic environ-
ment and the presence of proteolytic enzymes in the
gastrointestinal tract. In addition, peptides and
proteins exhibit a low permeability across the
gastrointestinal mucosa because they are hydrophilic
and have a high molecular weight (1-5). For orally ad-
ministered peptides and proteins to reach their site of
action, they must be able to resist chemical and en-
zymatic degradation in the gut lumen and then, after
penetration of the mucosal membranes, to escape first
- pass metabolism and clearance by the intestinal
mucosa and liver. Many peptide drugs can also be at-
 84 EUT. J. Drug Metab. Pharmacokinet; 1990, No.2

Table I

ROUTES OF DEGRADATION OF ORALL Y ADMINISTERED

PEPTIDES AND PROTEINS

I INTRALUMEN Gastrins and proteolytic enzymes of the pancreas

(trypsin, chymotrypsin, elastase, carboxypeptidases)

II BRUSH BORDER Absorbed pancreatic proteascs

MEMBRANES Aminopeptidases (oligopeplides)
Di, tri- and tetrapeptidases
Carboxypeptidases

III INTRACELLULAR Cytosolic peptidases (dipeptides > tripeptides)

Lysosomal proteases (oligopeptides and proteins)

Prot~in

tacted by peptidase activities in the blood circulation

(6-8).
Stomach

Co,.he"
medlote-d
transport
I All these handicaps encountered in the oral
delivery of peptides and proteins are not surprising
TO
ereee since the major function of the gastrointestinal tract is
the digestion of normal nutrition macromolecules (9).
Protein digestion starts in the stomach by the action of
Small
in16tine the endopeptidase pepsin (Fig. 1). This enzyme
provides as much as 10-30% of total protein digestion,
producing mainly polypeptides as hydrolysis products.
The majority of protein digestion, however, occurs in
"" the lumen of the small intestine by proteolytic enzymes
"" from pancreatic secretions, resulting in a mixture of
"" -, To
blood free amino acids and oligopeptides having 2 to 6 amino
" -, -, acid residues. This mixture is subsequently exposed to
the intestinal mucous membrane. The free amino acids
Fig.I: Schematic overview of the digestion and absorp- and the di - and tripeptides can be absorbed by active,
tion of dietary peptides and proteins. The shaded carrier - mediated transport systems. The tetra-, penta
area represents the intestinal brush border - and hexapeptides are much less absorbed intact, and
membrane. (Obtained form Gruber, Longer and are digested by brush border membrane peptidases
Robinson, ref. 9). (particularly aminopeptidases). Once inside the cell,

Table II

STRATEGIES FOR IMPROVED ORAL DELIVERY OF

PEPTIDE AND PROTEIN DRUGS

I Synthesisof stabilized and more lipophilic analogues

II Use of absorption enhancers and protease inhibitors
III Formulation approach: liposomes
pH - sensitive polyacrylicmating
azoaromatic polymer coating
biodegradable nanocapsules
bioadhesive microspheres
 1.Verhoefet al; Transport ofpeptide & protein dlugs
small peptides can be futher hydrolyzed to free amino
acids by intracellular peptidases. However, a small
percentage of these oligopeptides do escape this
hydrolysis process and do enter the blood intact. An
overview of proteases and peptidases involved in the
degradation of orally administered peptides and
proteins is given in Table 1.
In addition to the enzymatic barriers, the intestinal
absorption of peptide and protein drugs is severely
hampered by physical barriers as represented by the
mucous layer, the intestinal epithelial cell membranes,
and the tight junctions between the apical ends of the
epithelial cells (1, 9,10). The epithelial cells of the en-
tire intestine are covered by a mucous layer, consisting
of water, mucins (glycoproteins), electrolytes, proteins
and nucleic acids. The layer is bound to the apical cell
surface by the glycocalyx, a glycoprotein structure
which is covalently linked to the brush border
membrane. Due to the presense of sialic acid residues
and sulfate groups the mucins of the mucous layer are
negatively charged, which may result in electrostatic
binding or repulsion of the charged peptide and
protein molecules. The epithelial cell membranes are
composed of phospholipid bilayers, in which proteins
are embedded via their hydrophobic segments; these
bilayers represent very strong barriers for the transport
of very hydrophilic substances such as peptides and
proteins. Based upon the preceedingviews, it is evident
that peptide and protein drugs have little chance of
being absorbed intact from the intestine and that their
oral bioavailability will genarally be very low ( < 5%;
1,2). Nevertheless, oral delivery appears to be feasible
for certain peptides e.g. TRH (11) and arginine
vasopressin (12,13). Putative routes of intestinal
transport are the transcellular route (transport across
the epithelial cells) and the paracellular route
(transport between cells via the tight junctions and in-
tercellular space). Intestinal transport by passive
processes, probably both by the trans - and paracellular
route, has been reported for vasopressin and its
analogues (14,15). The transport of di - and tripeptides
is favoured by saturable peptide carrier - mediated
transport systems in the small intestine, which are dis-
tinct from the amino acid transport carriers (2,16).
Peptide drugs that have been shown to be transported
in a facilitated manner by carrier systems are TRH and
its analogues (1,16), dipeptide inhibitors of angioten-
sim converting enzyme (1,17) and p-lactam antibiotics
(structurally similar to tripeptides; 1,18). There is also
evidence for the intestinal transport of polypeptides
and proteins by fluid - phase of receptor - mediated
pinocytosis, e.g, for serum albumin and epidermal
growth factor (9,19).
85
STRATEGIES TO IMPROVE INTES-
TINAL TRANSPORT
In order to enhance the gastrointestinal absorption
of peptide drugs, several approaches have been inves-
tigate d in recent years (Table II). The first includes the
synthesis of peptide analogues with increased stability
to gastrointestinal enzymes and increased lipophilicity,
which may be accomplished by building in "unnatural"
amino acids (e.g, D - amino acids), by derivatization of
the NH2 - terminus, by cyclization, and by the use of C
- terminal blocking agents. This approach has resulted
in the development of orally active peptides, such as
TRH analogues (1), a behaviourally potent ACTH4-9
peptide (20), the antidiuretic compound DDAVP (12,
21, 22) and hypoglycemic dipeptide analogues (23).
The second strategy is dealing with the co - ad-
ministration of absorption enhancers and or protease
inhibitors (5, 10,24,25). There are five major types of
absorption enhancers: (a) chelators such as EDTA,
salicylates and N - acyl derivatives of collagen, (b) sur-
factants such as lauryl sulfate and polyoxyethylene - 9 -
lauryl ether, (c) bile salts such as glyco - and taurocho-
late, and derivatives such as taurodihydrofusidate, (d)
fatty acids such as oleic acid and capric acid, and their
derivatives such as acylcarnitines and mono - and
diglycerides, and (e) non - surfactants such as un-
saturated cyclic ureas. Most of the absorption
promoters are capable of increasing membrane
fluidity, either by creating disorders in the phos-
pholipid domain in the membrane or by facilitating the
leaching of proteins and lipids from the membrane
(24). They improve peptide and protein absorption
probably by one or combinations of several
mechanisms, which are summarized in Fig. 2. In-
t paracellular transport
t transcellular transport
• enzymatic degradatiOrf
• viscosity mucuslayer
-solubilization of drugs
Figure 2: Mechanisms of absorption enhancement.
86 Eur. J. DrugMetab. Pharmacokinet., 1990, No.2

creased oral bioavailability following co - administra-
tion with a variety of absorption enhancers has been
reported for several peptide drugs, including insulin
(26,27), the LHRH superagonist leuprolide (28), san-
dostatin (29), and calcitonin (30). The use of protease
inhibitors may also offer interesting possibilities for
oral delivery of peptide drugs (5, 25) and positive
results have been shown in the oral absorption of
tetragastrin (31), vasopressin (13), a nonapeptide in-
hibitor of renin (32), and insulin (33-35). It should be
noted here that the design and search for safe and ef-
fective absorption enhancing agents is presently be-
coming an important area of research: the trend ap-
pears now to be towards enhancers that are based on
natural constituents and that are biodegradable (De
Boer et aI., this Special Issue).
The third strategy to improve oral peptide delivery
is the formulation approach (Table II): the peptide
Cone.
[ng/mll
20
ban indicating S.E.M.
blol.dh.sl,.
( n. 3 )
15
10
5
3 4
time [hours]
drug is incorporated within a delivery system that is
designed to protect the peptide from contact with the
proteolytic enzymes in the intestinal lumen and to
release the peptide only upon reaching an area
favourable for its absorption. This may be due either
to low protease content or better permeability proper-
ties of the memhranes encountered. This approach has
particularly been investigated for the antidiabetic
polypeptide insulin, using water / oil / water - type
emulsions (36), entrapment in Iiposomes (37), encap-
sulation in soft gelatin capsules coated with pH - sen-
sitive polyacrylic polymers (38) or with polymers cross
- linked with azoaromatic groups (39), and nanocap-
sules of polyalkylcyanoacrylates (40, 41). Recently a
bioadhesive delivery system has been developed in our
Center, based on micro spheres of poly -2-
hydroxyethylmethacrylate and coated with muco - ad-
hesive polymers (42). This type of dosage form is aimed
to stick on the mucus linings of the intestinal track and
may exert a positive influence on peptide absorption in
a triplicate manner: retardation of gastrointestinal
transit, increase in local peptide concentration at the
site of adhesion / absorption, and decrease in diffusion
pathway from the delivery system to the absording
membrane. Loading the microspheres with the oc-
tapeptide DGAVP and using an in vitro model of rat
ileum, administration of coated microspheres resulted
in higher peptide levels in the serosal compartment as
compared to non - coated beads (Fig. 3). Present
research is now focussed on the potentials of these
5 bioadhesive microspheres in vivo.

Figure 3: In vitro absorption of DGAVP from muco - ad-

hesive and non - adhesive rnicrospheres in rat TRANSPORT ACROSS ALTERNATWE
ileum. The presented peptide levels were
measured in the serosalcompartment.(Data from ABSORPTIVE MUCOSAE
Lehr et al., ref. 42).
In the pharmaceutical research of peptide and
protein drugs alternative delivery routes with im-
TABLE III: Bioavailabilities of DEyE after rectaldelivery proved bioavailability are receiving much interest.
in rats These routes comprise the buccal, rectal, nasal and
pulmonary routes of administration. In comparison
with peroral delivery, these alternative mucosal routes
Delivery Adjuvant % Bioavailability
have the following advantages (5, 24): a) The existence
mode (mean)
of a rich vasculature, and avoidance or reduction of
first - pass elimination by the liver; b) Reduced
Infusion 0.5
presence of luminal proteolytic enzymes; c) The ab-
Infusion EDTA 0.6 sorptive epithelial membranes may be "leakier" than
Infusion MGK 14.6 the intestinal membranes, minimizing contact of the
Infusion EDTNMGK 25.7 peptides and proteins with the resident proteases; d)
Bolus 4.2 The peptide and protein drugs can be exposed to a
Bolus MGK 13.8 smaller surface area, and hence to a smaller fraction of
the total proteases; e) The compounds are subjected
Data from Van Hoogdalem et al. (ref. 55).
 J.Verhoefet al., Transport ofpeptide& protein drugs
to less dilution, thereby increasing the concentration
gradient for peptid and protein transport.
In should be emphasized that the epithelial cell
membranes of these alternative routes are also strong
physical barriers for the hydrophilic peptide and
protein drugs. Therefore, a great deal of research is
now being concentrated on the improvement of pep-
tide and protein transport across these alternative
mucosae, using strategies similar or comparable to
those summarized in Table II. A short overviewwill be
given of the present state of the art of peptide and
protein transport across non - peroral absorptive
mucosae, including recent results on peptide penetra-
tion through the skin.
Buccal transport
According to its natural function the buccal mucosa
is routinely exposed to a multitude of foreign com-
pounds, and is therefore likely to be less sensitive to ir-
reversible irritation or damage (by the dosage forms
and additives) than the other non - oral mucosae (43).
Several studies on the potential transport of peptide
drugs, with or without absorption enhancers, through
buccal epithelia have been reported, e.g. for TRH and
the LHRH analogue buserelin (43), a lauroyltripeptide
(44), the vasopressin fragment DGAVP (45), and in-
sulin (46, 47). Using effective absorption promoters
like glycocholate, laureth -9, and lauric acid in
propylene glycol, buccal insulin delivery in rats was
found to be one - fourth to one - third as effective as
intramuscularly injected insulin (47).
In terms of peptide permeability, the buccal
mucosa is genarally less efficient than the nasal and
rectal mucosae. The buccal route, however, is easy ac-
cessible and, by using appropriate muco - adhesive
dosage forms (43, 46, 48, 49), the buccal route of ad-
ministration offers opportunities for controlled pep-
tide delivery. A disadvantage maybe the obvious limits
imposed on the dimensions of a buccal adhesive gel or
patch and hence the dose that can be applied is limited
(50), making buccal delivery only suitable for highly
potent peptide drugs.
Rectal transport
The rectal route has obvious possibilities for pep-
tide and protein delivery, because its environment is
quite constant with respect to its pH, the amount and
viscosity of the rectal fluid, and its temperature (51).
There are also opportunities to reduce hepatic first-
87
pass elimination, particularly when the drug is ad-
ministered in the lower region of the rectum (52), and
to transport peptide and protein drugs to the lymphatic
sysem (53). Rectal drug delivery, however, is
hampered by a relatively low level of acceptance as a
route of drug administration (54). The feasibility of a
variety of absorption enhancers capable of increasing
the permeability of the rectal epithelium has been in-
vestigated for pentagastrin, gastrin, calcitonin and in-
sulin (review: ref. 4), and recently reported for the
neuropeptides DEyE, and DGAW, (55, 56) as wellas
for epidermal growth factor (57). By co - administra-
tion with sodium caprate and carboxymethyl cellulose
in rats, the rectal bioavailability of the latter polypep-
tide could be increased to approx. 70%. A major chal-
lenge in this area of research remains the search for
non - toxic and safe enhancing compounds (58).
Protease inhibitors can also be able to enhance rec-
tal peptide absorption (59), but data on the viabilityof
such compounds are very scarce. Nonetheless, studies
from our Center demonstrated that both absorption
enhancement and enzyme inhibition are prerequisites
for effective rectal delivery of labile peptide drugs like
DEyE (55). As depicted in Table II the metallo - pep-
tidase inhibitor EDTA did not have any effect on the
rectal bioavailabilityofDEyE although EDTA strong-
lyreduced the degradation of this peptide in the rectal
lumen (55); apparently, DEyE is not able to cross the
rectal mucosa if no absorption promoter is present.
The medium chain glyceride preparation MGK
markedly enhanced the biovailability of DEyE (ir-
respective of the deliveryrate; Table III), while co - ad-
ministration of MGK and EDTA produced a further
increase in bioavailability.
It is interestire to note that osmotic delivery sys-
tems (e.g. Osmet ) have the potentials for zero - order
and site - specific rectal drug administration in man.
These systems are very well tolerated, and also exhibit
rate - controlled deliveryof high - clearance drugs (52).
Thus these osmotic devices, in combination with the
use of absorption promoters and protease inhibitors,
may also have perspectives for rate - controlled rectal
administration of peptide and protein drugs.
Nasal transport
The nasal epithelial mucosa (classified into
respiratory and olfactory epithelial cells) is covered
with numerous microvilli, resulting in a large surface
area available for drug transport. For this reason,
together with the obvious advantages already men-
tioned, the nasal cavity is very suitable for peptide and
88 Eur. J. DTUg Metab. Pharmacokinet; 1990, No.2
protein delivery. At present a wide variety of such
drugs have been evaluated for intranasal absorption to
a level sufficient for displaying their biological effects;
these include gastrointestinal peptides (substance P,
secretin), pancreatic hormones (insulin, glucagon),
anterior pituitary polypeptides and proteins
(adrenocorticotropins, growth hormone), posterior
pituitary oligopeptides (oxytocin, vasopressin and
their analogues), hypothalamic releasing hormones
(TRH, LHRH, GHRH, somatostatin) and their
derivatives (e.g. the LHRH agonists buserelin,
histrelin and nafarelin), as well as enkephalins, cal-
citonin peptides and interferons (review: 60, 61). Very
recent evidence is available that polypeptides like
ACfH and calcitonin can be actively transported
across the nasal respiratory mucosa by endocytosis
(62). For nearly all peptide and protein drugs studied,
however, additives like absorption enhancers (60,61)
and/or protease inhibitors (5, 63, 64) are required in
order to achieve their systemic effects. For instance,
bile salts and fusidate derivatives have been shown to
increase nasal insulin transport by cooperation of dif-
ferent mechanisms, such as decreasing the viscosity of
the mucus layer, solubilization of the insulin molecules
in mixed micelles with these enhancers and subsequent
formation of reversed micelles within the nasal
epithelium, and by inhibiting enzymatic degradation
(61,65,66).
A major problem encountered in nasal drug
delivery may be the possible ciliotoxic and tissue
damaging effects of drugs and additives (67-69). The
,..
~
....
80
tZ
w powder dosage form with cellulose derivatives and
::> 60
@'
a:
u.
40
20
TIME (min)
Figure 4: Effect of STDHF (.), deoxycholate (.), and
laureth - 9 (~) on the ciliary beat frequency of
human adenoid tissue in vitro. Data ara the mean
± SD. Concentration of the enhancers used: 0.3%
(w.v.) (Data from Hermens et at., ref. 70).
nasal mucosa is covered by a mucus layer which is
transported to the throat by the nasal cilia, beating with
a frequency of approx. 15 beats/sec. This process,
known as the nasal mucociliary clearance, is an impor-
tant self - cleaning mechanism of the nose to protect
the body against inhaled toxic particles (dust, aller-
gens, bacteria), which should not be influenced by
nasal medication. In our laboratory it was found that
the well- known nasal absorption promoters laureth -
9 and deoxycholate cause a very rapid irreversible
ciliostasis (FIg. 4) The fusidate derivative STDHF at
concentrations of 0.3% and lower appears to be much
less ciliostatic than laureth - 9 and deoxycholate (Fig.
4) but more ciliotoxic than the trihydroxy bile sals
glyco-and taurocholate (70).
The nasal residence time is rather short and ex-
hibits high interindividual variability, ranging from 3 to
20 min in human volunteers (67). Therefore, the nasal
route commonly provides fast peak concentrations of
drug in the blood circulation. Thus nasal administra-
tion offers good opportunities for rapid and pulsatile
delivery of peptide and protein drugs. On the other
hand, the rapid nasal clearance of the dosage form may
be disadvantageous in obtaining reproducible absorp-
tion profiles (54). With the aim to prolong the nasal
residence time and to improve the reproducibility of
the absorption characteristics, impressive progress has
been reported using the formulation approach (Table
II). For example, nasal sprays may show prolonged
retention at the site ofapplication as compared to nasal
drops (71, 72) and the addition of viscous agents ap-
pears to delay the absorption of DDAVP from nasal
sprays (73). Bioadhesive polymer systems, in par-
ticular, may be expected to cause a break - through in
nasal drug delivery, provided that they do not strongly
interfere with the nasal ciliary movement. Nasal insulin
absorption can be enhanced and sustained by using a
polyacrylic acid (46); comparable effects have been ob-
served following nasal administration of insulin and
calcitonin in polyacrylic gel formulations (74). The
recently described bioadhesive microspheres (e.g, al-
bumin, starch) have a much longer clearance time from
the nose than spray and powder dosage forms (75),
enabling prolonged nasal absorption of insulin and,
most likely, also of other peptide and protein drugs
(76,n).
Pulmonary transport-
The pulmonary or inhalation route of drug ad-
ministration might also be favourable for the transport
 J. Verhoefet al; Transport ofpeptide & protein drugs
of peptide and protein drugs. It iswell accessible, there
is a large surface area of the alveolar epithelia, and
enormous vascularization of the subepithelial layer.
Already several years ago transport of insulin across
the respiratory mucosae has been reported to occur
following inhalation of aerosol dosage forms (78-80)
with bioavailability values up to 40% (80). Recent
studies have demonstrated that oligopeptides like
DDAVP and leuprolide can also pass over the
respiratory track to a high extent, and that the pul-
monary route for leuprolide delivery is superior to the
nasal and rectal routes of administration (81, 82).
Moreover, evidence is available that proteins like al-
bumin can be transported across the airway epithelia
by a low - affinity,receptor - mediated endocytotic sys-
tem (83). The usefulness of the pulmonary route in
drug delivery, however, seems to be limited by the risk
of infections in the airways and possible deposition of
aerosol particles.
Trandermal transport
Potential advantages of transdermal drug delivery
comprise the avoidance of hepatic first - pass effects,
the possibility of long - term application, maintenance
of a constant environment, ease to discontinue drug ad-
ministration, and excellent patient compliance. Disad-
vantages might be potential skin irritation and (local or
systemic) allergic reactions to drugs and excipients
(84). Although transdermal delivery of peptide and
protein drugs is ill-favoured by two major drawbacks,
extremely poor percutaneous transport and appreci-
...
=, II
..,
';:: 71
II: ..
U
ci ..
..."
-5
r ..
~ c
..
It
.
'. : .. , . . ,
:
• TI~E 'AtoL~sJ· .. .. .. I'
Figure 5: Cumulative penetration of ~-DE'YE through
human stratum corneum asa function of time. C,
Control; PG, stratum corneum pretreated with
propyleneglycol; Az, stratum corneum pretreated
with 10% Azone in propylene glycol. (Obtained
from Bodde, Verhoef and Ponee, ref. 85).
89
able intracutaneous breakdown (85), there seems to be
a future for this approach.
1'1 vitro percutaneous transport has been observed
for an a-MSH analogue across mouse but not rat skin
(86), for vasopressin across mouse and rat skin (87),
and for DEyE and DGAVP across human stratum cor-
neum (85, 88). Skin lipid fluidizers like Azone are
capable of enhancing the percutaneous penetration of
these peptides, as illustrated in Fig. 5 with DEyE data
from our labaratory. The degradation half - life of the
latter peptide, both in epidermis and dermis, was
surprisingly long as compared to that in human-plasma
(7, 85). Thus improvement of transdermal peptide
delivery will most likely occur by facilitating its per-
cutaneous transport, intra (epi-)dermal biotransfor-
mation being a problem of minor importance. In this
respect the recently developed iontophoretic delivery
devices appear to be very promising, particulary in pul-
satile-controlled peptide drug delivery. By applying
this new technique (poly) peptide penetration through
the skin can be impressively enhanced, as has been
reported for TRH, vasopressin and insulin in various
animal models (89-92) and for therapeutically relevant
doses of the LHRH analogue leuprolide in human
volunteers (93).
CONCLUSIONS
Peptide and protein drugs generally show high
potency and low toxicity, making these compounds
very attractive as therapeutic agents. On the other
hand, due to their unfavourable physio-chemical
properties, peptides and proteins are very poorly tran-
ported across biological membranes. These include
the epithelial mucosa of the intestinal tract as well as
other absorptive surfaces like the buccal, nasal and pul-
AZ monary epithelial mucosae. Many strategies can be
employed in order to facilitate peptide and protein
penetration through such physical and metabolic
membrane barriers. The synthesis of analogues with in-
creased metabolic stability, lipophilicity and receptor
PG
affinity offers good possibilities, but is obviously
restricted to (oligo) peptide drugs. The use of protease
inhibitors and particularly absorption enhancers is
another approach, being capable of decreasing the en-
II " II II
zymatic degradation and increasing the permeability of
the epithelial tissue, both for peptides and proteins. A
major problem encountered with the application of en-
hancers is their possible irritating and membrane
damaging effects, especially in the areas of the nasal,
rectal and pulmonary absorptive mucosae. Therefore,
research in this field will be directed towards the
search for non - destructive and safe absorption en-
hancing compounds from endogenous origin. And last
90 Eur. J. Drug Metab. Pharmacokinet; 1990, No.2
but not least, the recent advances in the development
of suitable dosage forms have very promising prospects
to improve peptide and protein drug delivery. This for-
mulation approach includes encapsulation in ap-
propriate polymer systems, bio (muco-) adhesive
microspheres, nanoparticles, etc. Using iontophoretic
delivery devices, even transdermal peptide delivery
seems to be possible in the near future.
It should be emphasized that pharmacokinetic -
pharmacodynamic relationships are very important is-
sues in the design of suitable drug delivery systems
(94). This also holds for peptide and protein drugs. For
instance, it is well known that the hypothalamic
luteinizing hormone - releasing hormone LHRH has
both stimulatory and inhibitory effects on pituitary
gonadotropin secretion, dependent on the input rate
of peptide administration (95). Pulsatile or time - pro-
grammed delivery of LHRH stimulates the release of
the gonadotropins LH and FSH from the pituitary
gland, and has shown to be therapeutically effective in
men and women with hypogonadotropic -
hypogonadism and in children with idiopathic delayed
puberty. On the other hand, continuous long - term ad-
ministration of LHRH and its analogues induces
desensitization of the pituitary LHRH receptors,
resulting in depressed pituitary secretions of LH and
FSH. The chronic continous mode of LHRH peptide
administration can be successfully applied in the treat-
ment of children with precocious puberty and patients
with prostate cancer. Thus these LHRH data clearly
illustrate that, in the design of appropriate dosage
forms for peptide and protein drugs, knowledge on the
mechanism of action and detailed clinical phar-
macological studies are essential, in order to establish
whether constant or pulsatile delivery of such com-
pounds is required for optimal pharmacotherapy.
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