2) United States Patent Nijkerk et al, US006613767B1 US 6,613,767 B1 *Sep. 2, 2003 a0) Patent No.: (45) Date of Patent: (54) STABLE AQUEOUS FOLINATE SOLUTION (75) Inventors: Alfred James Nikerk, Amsterdam (NL); Johanna M. P. Vermeer, Lisse (NL) (73) Assigaee: Pharmachemie B.Y., Haarlem (NL) (+). Notice: This patent issued on a continued pros- ecution application filed under 37 CFR 1.53), and is subject to the twenty year patent term provisions of 35 USC. 154(@\2). Subject to any disclaimer, the term of this patent is extended or adjusted under 35 ULS.C. 154(b) by 1258 days. (21) Appl. No.: 08/726,803 (22) Filed: Oct. 7, 1996 Related Application Data (63) Continuation in-pat of application No, PCTINL9Si00126, fled on Ape 4, 1905. (30) Foreign Application Priority Data Ape 5, 1994 (NL) 9400530 (51) Int. CL? AGIK 31/495 (52) US. Cl s14/249 (58) Field of Seareh 514249 66) References Cited U.S. PATENT DOCUMENTS 2,695,800 A 11/1954 Weideabeimer etal. a. 167165 S13423s A 7/1902 Mueller et al SA73ASS A * 121902 Hacger SM7005 A 9/1994 Mueller eta su20 FOREIGN PATENT DOCUMENTS EP mamas At 111988 EP 0203029 AL 11/1988 EP 0401895 12/1990 iP 0.667 159 8/1995 EP 0667159 A2 1995 Wo Woomidi60 9/1990 wo soo 9/1990, wo 95/26063 10/1995 OTHER PUBLIC ATIONS, Hagers Handbuch der Pharmazeutischen Praxis (1971), p. 30—"Stabilizers" Remington's Pharmaceutical Sciences (15° Ed.) Hoover et al,, Mack Pub. Co, Easton, PA. (1975) pp. 268-270 and 283-284," Massachusetts General Hospital, Dept. of Pharmacy, Dept of Nursing, Critical Care, “Tromethamine, Tris Buffer” btp:/www.mgh haruard .edu/pharmacy/ICU%2. guidelines! tromethamine.btm, Mar. 7, 2003." agers Handbook of the Pharmaceutical Practice, 1971, p. 358, (Germany) ae * cited by examiner Primary Examiner—Theodore J. Criares (74) Atorney, Agent, or Firm—Browdy and Neimark, PLLC 67) ABSTRACT stable aqueous solution of folinie acid of up to about 400 mgiml, which does not erystallize at refridgeration iperatures, is obtained by preparing an aqueous solution of sodium folinate 14 Claims, No DrawingsUS 6,613,767 BL 1 2 STABLE AQUEOUS FOLINATE SOLUTION ‘The above mentioned solution was found to be still stable after 3 months at refrigerator temperature between 2 and 8° C. (on average approximately 4° C), i. it had not erys- tallized out ‘This application is a continuation-in-part of PCT appl cation PCT/NI95\00126, filed Apr. 4, 1995, the entire ‘contents of which is hereby incosporated herein by refer- ‘ence. EXAMPLE 2 ‘The invention relates to a stable aqueous folinate solu- a tion, ‘The following injection Mid was prepared Folinie acid is a metabolite of folic acid and is the active form into which folic acid is converted in the body. This yp ‘conversion is inhibited by some cytostaties such as meth- ‘otrexate. In order to overcome this problem, folinic acid has Solum cae 030 me to be added. Folinic acid is further used in tbe case of folic ‘Sous chosue $00 ae acid deficiency, and it has a synergistic effect in combination Tes 1.21 mg with S-florouracl. The customary form in which flinic y< plumowitsogen ere 9 iam acid is added is caleium folinate Which is administered by infusion or injection, Caleium fotinateis readily soluble but it does not keep at room temperature and therefore cannot be ‘The thus obtained injection fluid was still stable after 3 stored for prolonged periods at room temperature. Al refrig- months at reigerator temperature. erator temperatures, where the flinate docs store well, 4 39 relatively concentrated solution isnot stable; at the normal EXAMPLE 3 refrigerator temperature of approximately 4° C., erystalliza~ tion takes place at a concentration of as Tittle as 15 mg/ml. EPO 401 895 provides folinate solutions with a relatively high concentration which are stable at refrigerator tempera~ 5 ture, These solutions contain folinate ions, caleium ions and ‘An injection fluid was prepared, having the following ‘components: 2 complexing agent for calcium. The complexing agent and Folscac (mim souem =i) 30 me calcium form a complex. These solutions may be stable at, ‘ime ons refrigerator temperatue ina concentration of as much as SO ‘Ses onée 40 tmgiml of folinate. The complexing agent used can be, ot 9 Tn prison che en ‘example, a sodium salt of ethylenediaminetetraacetic acid. ‘Water for injection to make up to 1 ithas been found that a sable aqueous solution of folinie acid! can be obiained. simply by preparing an aqueous - ; Solution of sodium foinate. Even ina high conectraton, up _‘TR¢obained injection Hid was ill stable afer 3. mons to about 40 mg of flinc acid perm ti stable inthe g Cltgetator temperature refrigerator, Le, does oo erysalize. "Te aqueous soltion of sodium folinate has a concentration of folinic acid in the EXAMPLE 4 range of about 15 to 400 mg/ml, preferably in the range of about 15 to 40 mg/ml or inthe range of about 20 10.400 Imgiml, and more preferably inthe cange of thot 25 10400 yo ig/ml. The solution is stable at a pil range of 40 0 100, specially from 6.0 to 100, and peteably about 80 to 9.0 ‘An injection uid was prepared, having the following ‘components: Preferably, the solution contains a stabilizer, sich as 250 me sodium citrate or sodium acetate, These salts further increase O30 me the stability of the solution. The present folinate solution Se ° also comprises an sotoniing agent such as sodium chloride, “* Wate bmjecton omae op “a Iris also advantageous to incorporate a pharmaceutically acceptable bulfer into the aqueous folinate solution aceord- ing to the invention. Such a pharmaceutically acceptable "This injection preparation was still stable after 3 months butler is preferably wis, a phosphate or a carbonate bufler, gy at effigerator temperature and is present in a concentration which is effective for obtaining the required pH. EXAMPLE 5 ‘Ths invention is further expsied in the follwing Ay igjeton vid was prepared, having the following examples. ‘components: EXAMPLE 1 ss ‘The following injection fluid was prepared Folin acd (oso salt) 250 me Sodiam site 035 Sosiam acetnte 038 lic aid (som fiat) 25 mg 6 Soin ehlorise 4.00 Seam ehlode 43 mg SaHLPO, LILO nase Sodium site 05 me NutiPo, 2100 i6t og ‘ater fr ijecton to make Up 10 1m Sodium hydtoxideynysrogen chloride 48 (OH 80) Water fr lection to make upto im “The pH had been set to a value of 75 by means of sodium 6s hydroxide. The purpose of the sodium chloride was to make This preparation was still stable after 3 months at reftig- the solution isotonic. erator temperature,US 6,613,767 BL 3 4 EXAMPLE 6 ‘An injection fuid was prepared, having the following ‘components: Siu Fain econ 26 male s ‘Tine period olin aid sun ml 750 mg Paces tsialy Glace 25.0 me Soe tte 050 ag ‘say fia aid 03% a 321 sscy Nba: Ot um hydoniehyoge ho = 10 besa gaan ak See eee a oe AeayNodomylfole 1 038 ait ‘Assay oie ait ae nt This injection preparation was still table after 3 months Taal sous ne 288 at reftigerator temperate ion pas 18. Min inl osm os EXAMPLE 7 tae pak I. Shelf Life Tests “a mena ot dec ‘Tests were carried out with sodium folinate bulk solutions ‘of 200 ml containing 25, 100, 250 and 400 mg sodium 20 TABLE 3 folinate/ml respectively as indicated in Table 1. Each bulk solution was prepared with a 2% excess of sodium folinate, Sodium olin nstion, 100 gin because during autoclaving ca. 2% folinic acid decomposes. “Tne priad ‘Subsquently, 2.15 ml of each formulated solution was used tare to fill 2R boities to the exclusion of ai. Afterwards, the 25 Parancies tsiatly 7 bottles were then autoclaved at 121° C. for 7 min. The —— aS bottles were thereafter stored at 40° C/75% RH (RH= = sll = slo relative humidity), The following analyses were carried out Passage yg ia saw lens) dens initially atthe beginning of the test and after two weeks: _betspae: mean range aie so Asay fain ci ise 9809 appearance Asay N(amino- 08% 09% pH Senup gmetad i ‘Seay Noy fle on 03% oxygen percentage in the headspace assy nm content of folie acid and decompesition products, ‘Assay oie id ade 01% ‘The results are summarized in the Tables 2-5 for the 5 Teal amount is 1 accelerated shelf life tests of the sodium folinate injection “> sitions peaks prepartion having content of sium flinnteof 25mg, Matai oo 0 TOO mim, 230 mpint and 400 mo respectively. The battles were sed at 40° C750 RH during the two week Taw Oe time period forthe shelf ie tess TABLE 1 Product 25 min 100 mgt 440 agit Sesion fine S57 g(2%) 223 g02%) S57 g(102) B92 g (10D Sain state “io 100 mg 100 mg 100 mg Sodio sloride 1000 me 1000 mg 1000 mg 1000 mg INNOWINHCL yp o pHTS £02 uptopH78202 yptopll78 +02 upto pil 78-02 ‘Water fr injection “Typo 2000 pe 200.0 ml upto 2000 ep 200. ml “The amount of lum flinte mat yet be cored forte content of salam flame in the raw mers ss TABLE 2 TABLE 4 ‘Sedum lisse Ineton, 25 mpi Sodium Folie Injection, 290 gil Tine pesiod Cy Tine pevod — Intaty 2 weeks a taitiany 2 weeks ‘Appestince Cleat, yellow soltion Clea, yellow sltion Appearance ‘Clea yellow-brown Chew, yellow brows mt 73 70 ‘scion lun Peestage oxygen in a9% (0-5) Lie(e=8 6S pit 73 72 headspoce pean ange 180% 3a Percentage oxygen imho: 2.6% (05) ad (a 4US 6,613,767 BL 5 6 lization took place. After storage for one month at 4° C., TABLE 4-continued crystallization was not observed in any of the four prepara: tions of Table 1 Sone Folie lesion 250 gin ‘The foregoing description of the specific embodiments 5 will so fully reveal the general nature ofthe invention that others ean, by applying current knowledge, readily modify fndor adapt for various applications such specific embod ments without departing Irom the generic concept, and, therefore, such adaptations and modifications should and are to intended to be comprehended within the meaning and range fiusaineo of equivaens of the disclosed embodiments. It 10 be Ky Nemes a8 ox understood that the phraseology’ or terminology employed Assay folic acid ade ade herein is for the purpose of description and not of limitation. ‘lan ts te ie cin forthe pr si trons owe cue gg _ At aco folnate solution for pharmaceutical appl- Ey cations consisting esentally of (@ sodium folinate, said soxtium fotinate being present in an effective amount at a concentration of up to 400 mg folinic acid per ml, Tn, mans “not deteated TABLE 5 Gi) a sabilizer selected from the group consisting of ‘sodium citrate, sodium acetate, and mixtures thereof, iam Folia ts Gi optionally, an isotonizing agent, and inc scot (iv) optionally, buffer selected from the group consisting ‘of tris, phosphate and carbonate bulers Sesame {sitially 2 weeks 25 2. The aqueous folinate solution according to claim 1, — Tianylowsors Cenyclowbore Wherein the pH ofthe solution i rom about 8.00 about 90 ion ion 3. The aqucous folinate solution according to elaim 1, fa "3 * wherein said solution contains said isotonizing agent Recxmmgcomacninteas —208n=8) nh =) 4, The aqueous folinate solution according to elim §, peer mean ange ane sree 30 Wherein said iotonizing agent is sodium ehforide see a ws coy 5m ns 5. The aqueous Folinte solution according to claim 1, Sasa wherein sti solution contains said buffer. Reap iofomybioiescid 1 um 6. The aqueous folinate solution aceording to claim 1, ‘Assay foie acid nase nd. ‘wherein said concentration of sodium Folinate is in the range ia 13% Hee of about 15 to 40 mg of folnie acid pee ml =e om 35 "7. The aqueous folate solution according to claim 1, peak nese cal poe wherein said concentration of sodium folinate is in the range Of about 20 to 400 mg of folic acid per ml “Tie cosa of fein mu ier ta he apeaed heowiialanomt "8, ‘The aqueous folinate solution aceording to claim 1, 10x Etat fh slaon ook morte ten wi he oer 3 wherein sald concentration of selium folate fein th range ub oon cee 40 of about 100 to 400 mg of folnic acid per ml. 9. The aqueous folinate solution according to claim 2, ‘The pH of all four formulations has been set before wherein suid concentration of sodium folinae is inthe range autoclaving t0 7,820.2, After autoclaving the pH of the of about 100 to 400 mg of folinic acid per ml sodium folinate injections of 25, 100, 250 and 400 mg/ml, 10. The aqueous folinate solution according to claim 3, respectively, decreased to 7.5, 7.1, 7.3 and 7.5, respectively. 45 wherein said concentration of sodium folinate isin the range ‘This pH reduction is probably due to the formation of the of about 100 to 400 mg of folinie acid per ml decomposition product N-(4-amino beazoyl glutaminic LL. The aquecus folinate solution according to claim 4, acid) in an amount of 03-0.5%. ‘wherein said concentration of sodium folinate is in the range Il appears from the results that the percentage of oxygen of about 100 t0 400 mg of folinie acid per ml in the headspace of the sodium folinate injection preparation, 12. The aqueous folinate solution according to claim 8, decreases to 1% (25 mg/ml formulation) or is no longer "wherein said concentration of sodium folinate is in the range detectable (the other three remaining formulations) alter of about 100 to 400 mg of folinie acid per ml storage for two weeks at 40° C/75% RH. 13. An aqueous folinate solution of sodium salts for ‘Surprisingly, the higher the content of sodium folinate in pharmaceutical applications, consisting of the formulation, the sinaller the decrease in the content after. "sodium folinate in a concentration of about 15 to 400 mg two weeks at 40° C/75% RH. In all four formulations there folinie acid per ml, was an inerease of 0.3-0.5% in the generation of the decomposition product N-(4-amino benzoyl glutaminic acid), This is probably the cause of the decrease in pH after 4 stabilizer selected from the group consisting of sodium citrate, sodium acetate, and mixtures thereof, two weeks storage at 40° C./75% RH. On the basis of the optionally, an isotonizing agent, present results it is concluded that the sodium folinate 60 optionally, a buffer selected from the group consisting of injection preparations stabilize with higher concentration of tris, phosphate and carbonate buffers, and ‘sodium folinate in the preparation optionally, a pH adjusting agent. 14. The aqueous folinate solution according to claim 13, UL Crystallization Tess werin sid concentration of sodium flinate isin he rage ‘The injection preparations shown in Table 1 were pre- 65 of about 100 to 400 mg of folinic acid per ml pared and stored at 4° C. to examine whether these prepa- rations remained stable or not, ie., whether oF not erystal- wees