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Europaisches Patentamt 

<D Q)JJ European Patent Office- <g> Publication number: 0 052 510 

Office europeen des brevets B 1 

® EUROPEAN PATENT SPECIFICATION 

® Date of publication of patent specification: 27.08 .86 ® Int. CI. 4 : A 61 K 9/50, A 61 K 9/52 
(8) Application number: 81305426.9 
® Date of filing: 17.11.81 



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<§) Microencapsulation of water soluble polypeptides. 



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Priority: 18.11.80 US 207864 



(§) Date of publication of application: 
26.05.82 Bulletin 82/21 



(§) Publication of the grant of the patent: 
27.08.86 Bulletin 86/35 



Designated Contracting States: 
AT BE CH DE FR GB IT LI LU NL SE 



References cited: 
DE-A-2051 580 
FR-A-2 400 904 
FR-A-2400 950 
US-A-3824227 
US-A-3826796 
US-A-3 887 699 
US-A-3 892723 
US-A-3 896105 
US-A-4 234 571 



(8) Proprietor: SYNTEX (U.S.A.) INC. 
3401 Hillview Avenue 
Palo Alto, California 94304 (US) 



@ Inventor: Kent, John Scott 
10120 Lockwood Drive 
Cupertino California 95014 (US) 
Inventor: Sanders, Lynda Mary 
765 San Antonio Road 65 
Palo Alto California 94303 (US) 
Inventor: Lewis, Danny Harvey 
312 Jackson Circle 
Gardendale Alabama 35071 (US) 
Inventor: Tice, Thomas Robert 
1305 Overland Drive 
Birmingham Alabama 35216 (US) 



Representative: Armitage, Ian Michael et al 
MEWBURN ELLIS & CO. 2/3 Cursitor Street 
London EC4A 1BQ(GB) 



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Note: Within nine months from the publication of the mention of the grant of the Eurbpeanpatent, any person may 
give notice to the European Patent Office of opposition to the European patent granted. Notice of opposition shall 
be filed in a written reasoned statement. It shall not be deemed to have been filed until the opposition fee has-been 
paid. (Art. 99(1) European patent convention). 



Courier Press, Leamington Spa, England. 



0 052 510 



Description 

This invention relates to pharmaceutical microcapsule compositions having sustained release 
characteristics where the active agent is a water-soluble polypeptide which is a luteinizing hormone/ 
5 releasing hormone, or analogue thereof, useful for affecting the reproduction function in mammals. 

There are a number of publications that disclose combinations of polymers and drugs designed to give 
sustained or delayed release of drugs. For example U.S. Patent 3,773,919 discloses controlled drug release 
compositions in which the core comprises a drug, stated to include water-soluble antibiotic polypeptides 
encapsulated in poly lactide/gly col ide copolymers as well as similar such polymers. 
10 Microencapsulation for sustained release of enzymes, hormones, vaccines and other bioiogicals is 
discussed in a paper by Chang, Thomas, J. Bioeng., Vol. 1, pp 25 — 32, 1976. Several examples of 
water-soluble protein encapsulations using polylactic acid are disclosed therein, particularly asparaginase 
and insulin compositions. 

Polylactic acid polymers, polylactide/glycolide copolymers and polyglycolic acid polymers and related 
is materials have been used for making surgical elements, incorporating a medicament and demonstrating 
slow release properties. See for example U.S. Patents 3,991,776; 4,118,470; 4,076,798. 

The invention relates to a pharmaceutical composition designed for sustained release of an effective 
amount of drug over an extended period of time prepared in microcapsule form wherein the composition 
comprises: 

20 at least one polypeptide which is a naturally occurring luteinizing hormone-releasing hormone 
(LH-RH), a synthetically prepared material of the same type or synthetically prepared analogues of naturally 
occurring LH-RH which act in some manner on the anterior pituitary gland to affect the release of luteinizing 
hormone (LH) and follicular stimulating hormone (FSH); 

optionally, at least one polymer hydrolysis modifying agent selected from organic acids, acid salts, 

25 neutral salts and basic salts; and a biocompatible, biodegradable encapsulating polymer which is a 
polylactide polymer, polyacetal polymer, polyorthoester polymer or polyorthocarbonate polymer. 

Said composition will release a daily amount of said polypeptide effective for maintaining an 
hormonally related condition over a predetermined period of time. The specific hormonally related 
condition bound to the use of said polypeptide is the control of fertility and physiological effects related 

30 thereto. 

The polymer hydrolysis modifying agents which may be optionally present in the composition may 
decrease or increase the rate of polymer hydrolysis. They have a low molecular weight and are non-toxic. 

The invention also relates to a process for preparing the above pharmaceutical composition, 
comprising: 

35 dispersing an aqueous solution containing the polypeptide, and optionally a polymer hydrolysis 
modifying agent, in a halogenated organic solvent containing said encapsulating polymer; 
adding to the dispersion a coacervation agent; and 
collecting the microcapsules from this solution. 

Naturally occurring LH-RH peptides are produced in the hypothalmic region of the brain and control 
40 the reproductive cycle of mammals by acting on the anterior pituitary gland to affect release of luteinizing 
hormone (LH) and follicular stimulating hormone (FSH) which in turn act on the gonads to stimulate the 
synthesis of steroid hormones and to stimulate gamete maturation. The pulsatile release of LH-RH thereby 
controls the reproductive cycle in mammals. Additionally, LH-RH has effects in placenta, in releasing HCG, 
and directly on the gonads. Agonist analogs of LH-RH are useful for the -control of fertility by two 
45 mechanisms of action. Low doses of LH-RH analogues can stimulate ovulation and are useful in the 
treatment of hypothalmic and ovulatory infertility. Additionally they can be used for hypogonadal 
conditions and impotence, and to stimulate spermatogenesis and androgen production in the male. 
Paradoxically, larger doses of highly potent and long-lasting analogues of LH-RH have an opposite effect 
and block ovulation in the female and suppress spermatogenesis in the mate. Related to these effects is a 
so suppression of normal circulating levels of sexual steroids of gonadal origin, including reduction in 
accessory organ weight in the male and female. In domestic animals this paradoxical effect promotes 
weight gain in a feed-tot situation, stimulates abortion in pregnant animals and in general, acts as a 
chemical sterilant. A full list of the paradoxical high dose effects is set out in<European Patent application 
EP— A— 21234. 

65 There is also the group of LH-RH analogues termed antagonists. These polypeptides have the 
paradoxical effect shown by LH-RH agonists but at low dose levels relative to naturally occurring LH-RH. 
Such compounds are to be included within the scope of this invention. 

The natural hormone releasing hormone LH-RH is a decapeptide comprised of naturally occurring 
amino acids (which have the L-configu ration except for the achiral amino acid glycine). Its sequence is as 

60 follows: 

(pyro)Glu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-Gly-NH 2 . 

Many analogues of this natural material have been studied. The beneficial effectiveness of these analogues 
65 has been varied. The most significant modification where agonists are concerned is obtained by changing 



2 



0 052 510 



the 6-position residue from Gly to a D-amino acid, for example, D-Ala, D-Leu, D-Phe or D-Trp. Antagonist 
activity can be best realized by substituting the naturally occurring 2-position His amino acid residue with 
with a D-amino acid residue. These analogues show increased activity relative to LH-RH. 

In addition to modifications position 6, increased agonist activity may be obtained by the following 
modifications: modifying position 10 to afford a nonapeptide as an alkyl-, cycloalkyl- or fluoroalkyl- amine, 
or by replacing Gly-NH 2 by an a-azaglycine amide; substituting N-methyl-leucine for leucine in position 7; 
replacing tryptophan in position 3 by 3-(1-naphthyi)-L-alanine; substituting the position 5 tyrosine residue 
with phenylalanine or 3-(1-pentafluorophenyl)-L-alanine; and the substitution at position 6 of unnatural 
D-amino acid residues containing two or more carbocyclic (or perhydroaryl) rings or a phenyl (or 
cyclohexyl) ring which is highly alkyl substituted. These specific compounds represent some of the more 
useful fertility affecting LH-RH type polypeptides which have been developed to date. This is not intended 
to be an exhaustive or exclusive list of all such compounds which which have been made or which can or 
may be made. They are simply set out to illustrate the type of compounds which are the subject of this 
invention. Any and all of them can be interchangeably substituted into the compositions of this invention. 

The compounds of specific interest herein are those from the last mentioned group wherein the 
6-position of the naturally occurring LH-RH material is replaced with a specific unnatural D-amino residue 
containing lipophilic carbocyclic residues, particularly residues containing two or more highly alkyl 
substituted carbocyclic aryi (or perhydroaryl) rings or a phenyl (or cyclohexyl) ring. These particular 
polypeptides are the subject of European Patent application EP — A — 21234, and are prepared in accordance 
with the procedures set forth therein. ' 

More specifically the polypeptides of particular interest in this invention are nonapeptides and 
decapeptides of the formula: 

(pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z (I) 

and the pharmaceutical^ acceptable salts there wherein: 

V is tryptophyl, phenylalanyl or 3-(1-naphthyl|-L-alanyl; 

W is tyrosyl, phenyalanyl or 3-{1-pentafluorophenyl)-L-aianyl; 
X is a D-amino acid residue 

O 

— N H — C H — C — 

CH 2 

I 

R 

wherein R is 

(a) a carbocyclic a ry (-containing radical selected from naphthyl, anthryl, fluorenyl, phenanthryl, 
biphenyl. benzhydryl and phenyl substituted with three or more -straight chain lower alkyl groups; or 

(b) a saturated carbocyclic radical selected from cyclohexyl substituted with three or more straight 
chain lower alkyl groups, perhydronaphthyl, perhydrobiphenylyl, perhydro-2,2-diphenylmethyl and 
adamantyl; 

Y is leucyl, Isoleucyl, nor-leucyl or N-methyMeucyl; 
Z is glycinamide or — NH — R,, wherein 

R, is lower alkyl, cycloalkyl, fluoro lower alkyl or 

0 

— NH — C — NH — R 2 

R 2 is hydrogen or lower alkyl. 

Preferred compounds of this invention are those wherein X is 3-(2-naphthyl)-D-alanyl or 3-(2,4,6- 
trimethylphenyl)-D-alanyl; Z is glycinamine; V is tryptophyl or phenylalanyl; W is tyrosyl and Y is leucyl or 
N-methyl-leucyl. 

Particularly preferred compounds are: 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 , 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-n-methyl-Leu-Arg-Pro-Gly-NH 2 , • 

(pyro)Glu-His-Phe-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 , 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2,4,6-trimethylphenyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 , 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-NHEt, 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-N-methyl-Leu-Arg-Pro-NHEt, 

and their pharmaceutical^ acceptable salts. 

Especially preferred is (pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 and its 

pharmaceutical^ acceptable salts. 



0 052 510 



As set forth above and for convenience in describing these compounds, the conventional abbreviation 
for the various amino acids are used as generally accepted in the peptide art as recommended by the 
IUPAC-IUB Commission on Biochemical Nomenclature, Biochemistry, 11, 1726 (1972) and represent the 
L-amino acids with the exception of the achiral amino acids in the 6-position designated by X. All peptide 
5 sequences mentioned herein are written according to the generally accepted convention whereby the 
N-terminal amino acid is on the left and the C-te'rminal amino acid is on the right. The abbreviation "Et" is 
monovalent ethane. 

As used herein, the term "pharmaceutically acceptable salts" refer to the salts that retain the desired 
biological activity of the parent compound and do not impart any undesired toxicoiogical effects. Examples 
io of such salts can be found in European Patent application EP — A — 21234, noted above. 

As used herein the term "lower alkyl" refers to a straight or branched chain saturated hydrocarbon 
group having from 1 to 4 carbon atoms such as, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, 
isobutyl, sec-butyl, and tert-butyl; the term "cycloalkyl group" refers to a cyclic saturated hydrocarbon 
group having from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; 
is the term "fluoro lower alkyl" refers to a lower alkyl group wherein one or more hydrogen atoms are 
replaced by fluorine, such as, for example, trifluoromethyl, pentafluoroethyl and 2,2,2-trifluoroethyl. 

As used herein "naphthyl" is inclusive of 1- and 2-naphthyi; "anthryl" is inclusive of 1-, 2- and 
9-anthryl; "fluoroenyl" is inclusive of 2-, 3-, 4-, and 9-fluoroenyl; "phenanthryl" is inclusive of 2-, 3- and 
9-phenanthryl; and "adamantyl" is inclusive of 1- and 2-adamantyl. 
20 As used herein the phrase "fertility affecting polypeptide" should be understood to mean any naturally 
occurring LH-RH polypeptide, synthetically prepared material of the same type or synthetically prepared 
analogues of naturally occurring LH-RH polypeptides which act in some manner on the anterior pituitary 
gland to affect the release of luteinizing hormone <LH) and follicular stimulating hormone (FSH); and in 
particular those polypeptides which inhibit ovulation or are useful for treating endometriosis in a female 
25 mammalian subject or are useful for treating benign prostatic hypertrophy and inhibiting spermatogenesis 
in a male mammalian subject. 

The compositions of this invention will contain the hormonally active polypeptides in varying amounts 
depending upon the effect desired. Treatment of infertility requires a low level of drug, while prevention of 
fertility and related effects requires a large dose relative to the activity of naturally occurring LH-RH. For the 
30 agonist fertility control it is expedient to prepare microcapsules which will release the drug at such a rate 
that the subject will receive between about 0.01 and 100 ug/kg body weight per-day, preferably between 0.1 
and 5.0 ug/kg body weight per day. 

The compositions of this invention are formulated to contain the polypeptide in an amount which may 
vary between 0.01 and 40.0 weight % of the polymer used for encapsulation. Preferably the peptide will be 
35 present in the amount between 0.1 to 10.0 weight %. 

The amount of drug placed in a particular formulation depends not only on the desired daily dose but 
also on the number of days that dose level is to be maintained. While this amount can be calculated 
empirically the actual dose delivered is a function of the degradation characteristics of the encapsulating 
polymer. Therefore the % weight of drug stated represent amounts which, when taken in conjunction with 
40 a particular polymer provide the desired release profile. 

Optionally, certain chemicals which affect the rate of polymer hydrolysis may be dissolved in the 
aqueous solution containing the polypeptide before it is encapsulated, by the polymer excipient. These 
chemicals are called polymer hydrolysis modifying agents. When present, these compounds may increase 
or decrease the rate at which the drug is released from the microcapsules. This effect is independent of a 
<5 particular polymer composition or size. 

Four types of chemicals may be used to realize this effect for example, organic acids, acidic neutral or 
basic salts. Low molecular weight mono and dicarboxylic acids such as acetic acid, tartaric acid, citric acid, 
gluconic acid, oxalic acid, ascorbic acid, succinic acid, their salts may be used. Basic salts may be, for 
example, ammonium sulfate, ammonium chloride, ammonium nitrate and sodium bisulphate. Neutral 
50 salts effective herein include metal halides such as, for example, sodium chloride, potassium chloride, 
sodium bromide, potassium bromide, calcium chloride and magnesium chloride. Basic salts include such 
salts as sodium carbonate, potassium carbonate, trisodium phosphate and tripotassium phosphate. Of 
these compounds it is most preferred to use either citric acid, sodium chloride or sodium carbonate. 
Combinations of these compounds will achieve the desired effect but the compositions described herein 
55 contain only one of these agents in a particular composition. 

When present the hydrolysis modifying agent will be added in an amount between 0.1 and 20% by 
weight of the polymer but preferably it will be present in the amount of 5 to 10%. 

The biocompatible, biodegradable encapsulating polymer which is used in this invention is a 
polylactide polymer, polyacetal polymer, polyorthoester polymer or polyorthocarbonate polymer. 
60 Of course, the polymer must be non-toxic to the host and must be of such composition that it is 
degradable by the body into metabolic products that have no deleterious or untoward effects on the body. 
The polymer must also be capable of forming microcapsules containing water-soluble drugs. 

A number of particular polymers have been developed which meet these criteria. Various 
combinations of alpha hydroxycarboxylic acids and certain lactones can be condensed to form such 
55 polymers, particularly lactic acid and glycolic acid or combinations thereof. See, for example U.S. Patent 



4 



0 052 510 

No. 3773,919. Similar biocompatible polymers based on glycolic acid and glycerol and the like also are . 
known. See U.S. Patents 3,991,776; 4,076,779 and 4,118,470 for examples of such compositions. Several 
new biocompatible, biodegradable polymers derived from polyorthoesters and polyorthocarbonates also 
may be effectively used an encapsulating excipients in the practice of this invention. These latter polymers 

s are described in U.S. Patents 4,093,709 and 4,138,344. There are also known polyacetals and 
polyorthoesters useful for this purpose as described in Polymer Letters 75, 293 (1980). This list is not 
intended to be exhaustive of the particular polymers which are compatible with the scope of this invention 
but merely sets out examples to illustrate the polymers which may be used. 

One preferred group of polymer excipients are the orthoester and orthocarbonate polymers having a 

w repeating unit comprising a hydrocarbon radical and a symmetrical dioxycarbon unit of the general 
formula: 



R-i — O — C — l 




wherein R, is a multivalent hydrocarbon radical, R 2 and R 3 are hydrocarbon radicals with at least one of R a 
20 or R 3 bonded to the dioxycarbon through the oxygen linkage, and which polymers are synthesized by 
reacting a polyol with an orthoester or orthocarbonate. A full and complete description of the exact 
compositions, preparation, and properties of these polymers can be found in U.S. Patents 4,093,709 and 
4,138,344. 

Also preferred are those polymers based on the condensation of di vinyl ethers and polyols. These 

25 compounds are prepared by reacting polyol with a diketene acetal to form the polyacetal. A more detailed 
description and discussion of these polymers can be found in the journal, Polymer Letters, J. Heller, et al, 
18, 293 (1980). Of similar interest are those polyorthoesters prepared by a modification of the synthesis 
used to prepare the above polyacetals. These polymers are comprised of diketene acetal-diol condensates. 
For example, the diketene acetal 3,9-bis-(methylene)-2,4,-8,10-tetraoxaspiro[5,5lundecane can be 

30 condensed with 1 ,6-hexanediol to give a polyorthoester polymer which has degradation properties in vivo 
which make its use in the compositions of this invention desirable. Further preparation techniques and 
polymer characteristics for these compounds can be found in U.S. Patent Nos. 4,093,709; 4,131,648; 
4,138,344; and 4,180,646. 

Most preferred herein are those polymers derived from the condensation of alpha hydroxycarboxylic 

35 acids and related lactones. The most preferred polymer excipients herein are derived from an alpha 
hydroxy acid, particularly lactic acid, glycolic acid or a mixture of the two. 

The alpha hydroxy acid units from which the preferred excipients are prepared may be the optically 
active (f> and L-) forms or optically inactive (DL-, racemic) forms. For example, lactic acid, whether it is the 
principle polymer component or the comonomer component, can be present as D-lactic acid, L-lactic acid 

40 or DL-lactic acid. 

Other comonomers, for example certain C3 to C18 carboxylic acids and certain lactones, can be used in 
the preparation of preferred polymers. Illustrative of such compounds are 3-propiolactone, 
tetramethy (glycol ide, b-butyrolactone, 4-butyrolactone, pivalolactone, and intermolecular eyelid asters of 
a-hydroxy butyric acid, a-hydroxyisobutyric acid, a-hydroxy valeric acid, a-hydroxyiso valeric acid, a- 

45 hydroxy caproic acid, a-hydroxy-a-ethylbutyric acid, a-hydroxyiso pea proic acid, a-hydroxy-3-methylvaleric 
acid, a-hydroxy hepta no ic acid, a-hydroxy octanoic acid, a-hydroxydecanoic acid, a-hydroxy my ristic acid, 
a-hydroxystearic acid, and a-hydroxy lignoceric acid. 

Any of these compounds may be used a comonomer in the preparation of acceptable polymers. 
3-butyrolactone can be used as the sole monomer or as the principle monomer along with any of the 

50 comonomers recited above. However it is most preferred to use lactic acid as the sole monomer or lactic 
acid as the principle monomer with glycolic acid as the comonomer. 

The term pofyiactide is used to designate the general glass of polymers which can be prepared from 
one or more of the preferred monomers listed above and includes those instances where a single alpha 
hydroxy acid or lactone is the only monomer in the polymer. For the most preferred polymers, those 

55 wherein the excipients are prepared solely from the lactic acid monomer or where lactic acid is the principle 
monomer and glycolic acid is the comonomer are termed poly(lactide-co-glycolide) copolymers. 

The combinations of prefered monomer and comonomer which can be prepared are numerous but the 
most effective excipients are those polymers prepared from lactic acid alone or lactic acid and glycolic acid 
wherein the glycolic acid is present as a comonomer in a molar ratio of 100:0 to 40:60. It is most preferred 
60 to use a poly(tactide-co-glycolide) copolymer having a molar ratio between about 75:25 and "50:50. 

Poly(lactide-co-glycolide) polymers may range in size from 20,000 to 100,000 in molecular weight, 
stated as an average. The molecular weight of a particular copolymer is independent of its monomeric 
makeup. For example, a 50:50 copolymer can have a molecular weight which falls anywhere within this 
range. Therefore polymers can be varied both as to their monomer composition and as well as their 
55 molecular weight and be within the scope of this invention. 



5 



0 052 510 



For the purposes of this invention the relative molecular weight of a particular polymer vis-a-vis a 
second polymer is stated in terms of its inherent viscosity in a particular solvent and at a particular 
temperature. The viscosity of a particular polymer is measured in a capillary viscometer using chloroform 
or hexafluoroisopropanol at 30 C C. The results are stated in terms of deciliters/g (dl/g). There is a direct 
s correlation between inherent viscosity and molecular weight. 

A method for the preparation of polylactide polymers can be found in U.S. Patent 3,773,919 and 
reference is made thereto for the preparation of the such polymers. 

Preparation of the microcapsules using any combination of the various peptides, polymer hydrolysis 
modifying agents or encapsulating polymer excipients noted above parallels the basic technique set out in 
io U.S. Patent 3,773,919. A full description of the procedure used herein can be found in that document. 

In brief, the procedure involves dissolving the polymer in an halogenated hydrocarbon solvent, 
dispersing the aqueous drug solution in this polymer-solvent solution, and adding some agent which is 
soluble in the halogenated hydrocarbon solvent but is a non-solvent for the encapsulating excipient. The 
addition of the non-solvent, called a coacervation agent, causes the excipient to precipitate out of the 
is halogenated hydrocarbon solvent onto the dispersed water droplets, thereby encapsulating the 
polypeptide. For example, a poly(lactide-co-glycolide) is dissolved in methylene chloride. An aqueous 
solution of polypeptide is then stirred into the solvent-polymer solution to form a water-in-oil emulsion. A 
second solvent-miscible material such as a silicone oil, is added slowly with stirring to precipitate the 
excipient which coats the dispersed water droplets to give microcapsules. 
20 Halogenated organic solvents which may be used are most of the C1 to C4 halogenated alkanes such 
as, for example, methylene chloride, ethylene dichloride, ethylene chloride and 2,2,2-trichloroethane. 

Coacervation agents may be any solvent miscible polymeric, mineral oil or vegetable oil compounds 
which are non-solvents for the encapsulating polymers. There may be used, for example, silicone oil, 
peanut oil, soybean oil, corn oil, cotton seed oil, coconut oil, linseed oil, and mineral oils. 
25 After being formed, the microcapsules are washed and hardened with an alkane organic solvent, 
washed with water, washed with an aqueous non-ionic surfactant solution, and then dried at room 
temperature under vacuum. 

Microcapsules may range in diameter from about 1 to 500 urn, depending upon the techniques 
employed. For this invention it is preferred to have the microcapsule diameter be between 5 and 200 urn. 
30 The prepared microcapsules may be administered to a subject by any means or route desired. 
However the most effacious route is parenteral administration by injection, most preferably 
subcutaneously or intramuscularly. 

If the capsules are to be administered by injection they may first be suspended in some non-toxic 
suspending vehicle. The exact make up of these injectable microcapsule suspensions will depend upon the 
36 amount of drug to be administered, the suspending capacity of the suspending agent and on the volume of 
solution which can be injected at a particular site or in a particular subject. 

The compositions of this invention exhibit sustained release of the encapsulated compounds over 
extended periods of time. This time period may range from one month to 3 years depending on the 
composition of the encapsulating excipient, its molecular weight, the diameter of the capsule, and the 
40 presence of a polymer hydrolysis modifying agent in the core. Preferably the release time will be about 1 to 
24 months. 

The following examples illustrate the compositions and processes of this invention. 
Example I 

45 This example describes the procedure for preparing a microcapsules composition wherein the 
polypeptide is 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 , 

so (D-Nal(2) 6 LH-RH) present in an amount of 1.4% by weight, no polymer hydrolysis modifying agent is 
present, and the excipient is a 50:50% molar ratio poly(lactide-co-glycotide) copolymer having an inherent 
viscosity in hexafluoroisopropanol of 0.38 dl/g at 30°C. 

Excipient, 4 g, were dissolved in 196 g of methylene chloride. This solution was placed in a 300 ml resin 
kettle equipped with a true-bore stirrer having a 63 mm. Teflon (Trade Mark) turbine impeller driven by a 

55 Fisher "Steadi-Speed" (Trade Mark) motor. In a 1-drarn glass vial was dissolved 0.0571 g of polypeptide in 
1.34 g of deionized water. This solution was added to the resin kettle. During this addition, the dilute 
polymer solution was stirred at 3200 RPM to form a water-in-oil emulsion. With continued stirring at that 
rate, 80 ml of silicone oil was added at the rate of 4.0 ml/min by means of a peristaltic pump. The silicone oil 
caused the polymer to phase separate, and deposit as droplets of solvent-swollen polymer onto the surface 

bo of the water-poly peptide microdroplets. These solvent-swollen polymer droplets then coalesced to form a 
continuous film around the water-poly peptide microdroplets. The microcapsules were then hardened by 
pouring the contents of the resin kettle into a beaker containing 2000 ml of heptane. This mixture was 
stirred at 1000 RPM for 30 minutes with a stainless-steel impeller. The heptane-methylene chloride-silicone 
oil solution was removed by filtering the solution, employing a Buchner funnel and Whatman 041 filter 

65 paper. The microcapsules were then washed repeatedly with 100-ml aliquots of heptane to insure complete 



6 



0 052 510 



removal of the silicone oil. The microcapsules were then washed with deionized water followed by a wash 
with a 1% aqueous solution of Tween 20 (Trade Mark), and dried at room temperature under vacuum. 
Microcapsules obtained from this preparation were determined to have diameters ranging in size from 10 
to 40 prn. 

5 The polypeptide containing microcapsules, whose preparation is described in the above paragraph, 
were suspended in a suspending vehicle and administered as a single subcutaneous injection to female 
Sprague-Dawley rats and female rhesus monkeys. The length of estrous suppression was calculated 
against the percentage of animals showing suppression. 

The results of the monkey study are given in Table I below. Each data line represents one subject. The 

w injected dose was as stated in the Table. Microcapsules were prepared as stated in Example I using that 
LH-RH analogue and a 50:50% molar ratio copolymer (P LA: PGA} having an inherent viscosity of 0.38 dl/g in 
hexafluoroisopropanol at 30°C at a 1.4% peptide to polymer ratio. The microcapsule's diameter ranged 
from 10 to 40 um. 

is TABLE I ^ 

Effect of D-Nal(2) 6 LHRH released from PLA:PGA microspheres on ovulation in rhesus monkeys 



Animal 
No. 


Dose 


Before 


Intermenstrual interval 

During After treatment 


1 




25 


30 


28 


2 




28 


27 


26, 29 


3 


1 mg 
D-Nal{2) 6 


30 


67 


27 


4 


1 mg 
D-Nal(2) 6 


24 


83 


27 



A single 300 ug dose of D-Nal(2) 6 LH-RH micro-encapsulated at 1.4% peptide to polymer with a 50:50% 
molar ratio poly(lactide-co-glycolide) having a diameter ranging in size from 10—^40 um (inherent viscosity 
in hexafluoroisopropanol-0.38 dl/g) which had been suspended in a suspending agent (composition given 
35 in Example III) was injected subcutaneously in 10 mature female Sprague-Dawley rats. Estrous was 
determined by daily vaginal smear analysis. All rats showed estrous suppression through day 24 post 
dosing. At day 25, 40% showed estrous. By day 27 estrous was observed in all animals. 

Example II 

40 Table II sets out several examples of polypeptide containing microcapsules wherein the following 
parameters were varied: lactide-glycolide mole ratio; molecular weight, stated as inherent viscosity; stir 
rate; addition rate of silicone oil; and the amount of silicone oil added. The polypeptide encapsulated here 
is the same as set out in Example I. The preparation techniques described in Example I were used to 
prepare these materials, except as note for the stirring rates and silicone oil addition rates. 

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Example HI 

The following describes a formulation for parenteral injection of polypeptide-containing microcapsules 
prepared according to the methods disclosed herein. 
Microcapsules containing the polypeptide 



(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH a 

in a concentration of 1.0% by weight and wherein the excipient polymer was poly{lactide-co-glycolide) 
having a molar ratio of 50:50% and an inherent viscosity of 0.38 dl/g in hexafluoroisopropanol at 30°C were 
10 suspended in the following solution: 

Na CMC 0.5% 

NaCI 0.8% 

15 

Benzyl alcohol 0.9% 

Tween 80 (Trade Mark) 0.1% 

20 Purified water q.s. 100% 

For example, 330 mg of microcapsules were suspended in 5.5 ml to provide an injectable dose of 300 
ug of peptide per 0.5 ml of injectable suspension. 

25 Claims for the Contracting States: BE, CH, DE, FR, GB, IT, U, LU, NL, SE 

1 . A pharmaceutical composition designed for sustained release of an effective amount of drug over an 
extended period of time prepared in microcapsule form wherein the composition comprises: 

at least one polypeptide which is a naturally occurring luteinizing hormone-releasing hormone 
30 (LH-RH), a synthetically prepared material of the same type or synthetically prepared analogues of naturally 
occurring LH-RH which act in some manner on the anterior pituitary gland to affect the release of luteinizing 
hormone (LH) and follicular stimulating hormone (FSH); 

optionally, at least one polymer hydrolysis modifying agent selected from organic acids, acid salts, 
neutral salts and basic salts; and 
35 a biocompatible, biodegradable encapsulating polymer which is a polylactide polymer, polyacetal 
polymer, polyorthoester polymer or polyorthocarbonate polymer. 

2. A composition of claim 1 wherein said polypeptide is a nonapeptide or a decapeptide analogue of 
LH-RH having the formula 

40 (pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z 

and the pharmaceutical^ acceptable salts thereof wherein: 
V is tryptophyl, phenylalanyl or 3-{1-naphthyl)-L-alanyl; 
W is tyrosyl, phenylalanyl or 3-{1-pentafluorophenyl)-L-alanyl; 
45 X is a D-amino acid residue 

O 

— IMH — CH — C — 



CH 2 



wherein R is 

55 (a) a carbocyclic aryl-containing radical selected from naphthyl, anthryl, fluorenyl, phenylanthryl, 
biphenylyl, benzhydryl and phenyl substituted with three or more straight chain C,^ alkyl groups; or 
(b) a saturated carbocyclic radical selected from cyclohexyl substituted with three or more straight 
chain C w alkyl groups, perhydronaphthyl, perhydro- biphenylyl, perhydro-2,2-dipbenylmethyl and 
adamantyl; 

60 Y is leucyl, isoleucyl, nor-leucyl or N-methylleucyl; 

2 is glycinamide or — NH — R v wherein R, is Ci- A alkyl, C 3 - 6 cycloalkyl, fluoro C,_ 4 alkyl or 

O 

li 

65 — NH — C — NH — R 2 



9 



0 052 510 



R 2 is hydrogen or d- 4 alkyl. 

3. A composition of claim 2 having a polymer which is a poly(lactide-co-glycolide) copolymer wherein 
the copolymer comprises lactide-glycolide in a molar ratio of between 100:0 and 40:60; and wherein the 
copolymer has an average molecular weight between about 20,000 and 100,000. 
5 4. A composition of claim 3 wherein said polypeptide is present in an amount of between 0.01 and 40.0 
weight % of the polymer; and said hydrolysis modifying agent is present in an amount of between 1 and 15 
weight % of the polymer. 

5. A composition of claim 3 or claim 4 having a polypeptide wherein: 

V is tryptophyl or phenylalanyl; 
70 W is tyrosyl; 

X is 3-(2-naphthyl)-D-alanyl or 3-{2,4,6-trimethylphenyl)-D-alanyl; 

Y is leucyl or N-methyl-leucyl; and 
Z is glycinamide or NHEt; 

said hydrolysis modifying agent is citric acid, ammonium chloride, sodium chloride or sodium 
is carbonate; and 

said polymer comprises lactide-co-glycolide in a molar ratio of between 75:25 and 50:50. 

6. A composition of claim 5 wherein said polypeptide is present in an amount of 0.1 to 10.0 weight %; 
said hydrolysis modifying agent is present in an amount of 5 to 10 weight %; and 

said polymer comprises lactide-co-glycolide in a molar ratio of 50:50. 
20 7. a composition of any one of claims 1 to 6 wherein said polypeptide is 

{pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 

or a pharmaceutical^ acceptable acid salt thereof. 
25 8. A composition according to claim 1, wherein the polypeptide is an analogue of natural LH-RH, in 
which modification comprises the 6- posit ion residue changed from Gly to a D-amino acid. 

9. A composition according to claim 8, wherein the D-amino acid is D-Ala, D-Leu, D-Phe, or D-Trp. 

10. A composition according to claim 9, wherein the D-amino acid is D-Leu. 

11. A composition according to claim 9, wherein the D-amino acid is D-Trp. 

30 12. A composition according to any one of claims 8 to 11, wherein the 10-position is modified to afford 
a nonapepttde as an alkyl-, cycloalkyl- or fluoroalkyl-amine. 

13. A composition according to any one of claims 8 to 11, wherein the Gly-NH 2 is replaced by an 
a-azaglycine amide. 

14. A composition according to any one of claims 8 to 11, wherein N-methyl-leucine is substituted for 
35 leucine in position 7. 

15. A composition of any one of claims 7 to 14 wherein the polymer is a poly(lactide-co-glycolide) 
copolymer. 

16. A composition of claim 15 wherein the copolymer comprises lactide-glycolide in a molar ratio of 
between 100:0 and 40:60. 

40 17. a composition of any one of claims 7 to 16 wherein said polypeptide is present in an amount of 
between 0.01 and 40.0 weight % of the polymer. 

18. A composition of any one of claims 7 to 17 wherein said hydrolysis modifying agent is present in an 
amount of between 0.1 and 20 weight %. 

19. A composition of any one of claims 7 to 18 wherein said polypeptide is present in an amount of 0.1 
*s to 10.0 weight %. 

20. A composition of any one of claims 7 to 19 wherein said polymer comprises lactide-glycolide in a 
molar ratio of from 75:25 to 40:60. 

21. A composition of any one of the preceding claims in the form of injectable particles ranging in size 
from about 1 to 500 um. 

so 22. A composition of any of claims 1 to 21 which are dispersed in a pharmaceutical^ acceptable carrier 
suitable for parenteral administration. 

23. A process for preparing a composition of any one of the preceding claims comprising: 
dispersing an aqueous solution containing the polypeptide, and optionally a polymer hydrolysis 
modifying agent, in a halogenated organic solvent containing said encapsulating polymer; 
55 adding to the dispersion a coacervation agent; and 
collecting the microcapsules from this solution. 

Claims for the Contracting State: AT 

60 1 . A process for preparing a pharmaceutical composition designed for sustained release of an effective 
amount of drug over an extended period of time prepared in microcapsule form; characterised in that the 
process comprises: 

dispersing an aqueous solution containing a polypeptide which is a naturally occurring luteinizing 
hormone-releasing hormone ( LH-RH}, a synthetically prepared material of the same type or synthetically 
65 prepared analogues of naturally occurring LH-RH which act in some manner on the anterior pituitary gland 



10 



0 052 510 



to affect the release of luteinizing hormone (LH) and follicular stimulating hormone (FSH), and optionally a 
polymer hydrolysis modifying agent which is an organic acid, acid salt neutral salt or basic salt, in a 
halogenated organic solvent containing a biocompatible, biodegradable encapsulating polymer which is a 
polylactide polymer, polyacetal polymer, polyorthoester polymer or polyorthocarbonate polymer; 
5 adding to the dispersion a coacervation agent; and 
collecting the microcapsules from this solution. 

2. A process of claim 1 wherein said polypeptide is a nonapeptide or a decapeptide analogue of LH-RH 
having the formula 

io (pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z 

and the pharmaceutical ly acceptable salts thereof wherein: 
V is tryptophyl, phenyialanyl or 3-(1-naphthyl)-L-alanyl; 
W is tyrosyl, phenyialanyl or 3-(1-pentafluorophenyl)-L-alanyl; 
is X is a D-amino acid residue 

0 

— NH — CH — C — 

20 i 

CH 2 



25 wherein R is 

(a) a carbocyclic aryl-containing radical selected from naphthyl, anthryt, fluorenyl, phenylanthryl, 
biphenylyl, benzhydryl and phenyl substituted with three or more straight chain C,- 4 alkyl groups; or 

(b) a saturated carbocyclic radical selected from cyclohexyl substituted with three or more straight 
chain C w alkyl groups, perhydronaphthyl, perhydrobiphenylyl, perhydro-2,2-diphenylmethyl and 

30 adamantyl; 

Y is leucyl, isoleucyl, nor-leucyl or N-methyl-leucyl; 

2 is glycinamide or — NH — R„ wherein R n is Cw alkyl, C 3 _ fi cycioalkyl* fluoro C t . 4 alkyl or 

0 

35 ;] 

— NH — C — NH — R 2 



R 2 is hydrogen or d_« alkyl. 
40 3. A process of claim 2 using a polymer which is a poly(lactide-co-glycolide) copolymer wherein the 
copolymer comprises lactide-glycolide in a molar ratio of between 100:0 and 40:60; and wherein the 
copolymer has an average molecular weight between about 20,000 and 100,000. 

4. A process of claim 3 wherein said polypeptide is present in an amount of between 0.01 and 40.0 
weight % of the polymer; and said hydrolysis modifying agent is present in an amount of between 1 and 15 

45 weight % of the polymer. 

5. A process of claim 3 or claim 4 using a polypeptide wherein: 
V is tryptophyl or phenyialanyl; 

W is tyrosyl; 

X is 3-<2-naphthyl)-D-alanyl or 3-(2,4,6-trimethylphenyl>-D-alanyI; 
50 Y is leucyl or N-methyl-leucyl; and 
2 is glycinamide or NHEt; 

said hydrolysis modifying agent is citric acid, ammonium chloride, sodium chloride or sodium 
carbonate; and 

said polymer comprises lactide-co-glycolide in a molar ratio of between 75:25 and 50:50. 
55 6. A process of claim 5 wherein said polypeptide is present in an amount of 0.1 to 10.0 weight %; 
said hydrolysis modifying agent is present in an amount of 5 to 10 weight %; and 
said polymer comprises lactide-co-glycolide in a molar ratio of 50:50. 

7. A process of any one of claims 1 to 6 wherein said polypeptide is 

60 (pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 

or a pharmaceutical^ acceptable acid salt thereof. 

8. A process according to claim 1, wherein the polypeptide is an analogue of natural LH-RH, in which 
modification comprises the 6-position residue changed from Gly to a D-amino acid. 

65 9. A process according to claim 8, wherein the D-amino acid is D-Ala, D-Leu, D-Phe, or D-Trp. 



11 



0 052 510 



10. A process according to claim 9, wherein the D-amino acid is D-Leu. 

11. A process according to claim 9, wherem the D-amino acid is D-Trp. 

12. A process according to any one of claims 8 to 11, wherein the 10-position is modified to afford a 
nonapeptide as an alkyl-, cycloalkyl- or fluoroalkyl-amine. 

13. A process according to any one of claims 8 to 11 ( wherein the Gly-NH 2 is replaced by an 
a-azaglycine amide. 

14. A process according to any one of claims 8 to 11, wherein N-methyl-leucine is substituted for 
leucine in position 7. 

15. A process of any one of claims 7 to 14 wherein the polymer is a poly(lactide-co-glycolide) 
copolymer. 

16. A process according to claim 15 wherein the copolymer comprises lactide-glycolide in a molar ratio 
of between 100:0 and 40:60. 

17. A process of any one of claims 7 to 16 wherein said polypeptide is present in an amount of between 
0.01 and 40.0 weight % of the polymer. 

18. A process of any one of claims 7 to 17 wherein said hydrolysis modifying agent is present in an 
amount of between 0.1 and 20 weight %. 

19. A process of any one of claims 7 to 18 wherein said polypeptide is present in an amount of 0.1 to 
10.0 weight %. 

20. A process of any one of claims 7 to 19 wherein said polymer comprises lactide-glycolide in a molar 
ratio of from 75:25 to 40:60. 

21. A process of any one of the preceding claims wherein the product is in the form of injectable 
particles ranging in size from about 1 to 500 urn. 

22. A process of any of claims 1 to 21 wherein the product composition is dispersed in a 
pharmaceutically acceptable carrier suitable for parenteral administration. 

Patentanspruche fur die Vertragsstaaten: BE, CH, DE, FR, GB, IT, II, LU, NL und SE 

1. Pharmazeutische Zusammensetzung fur die verzogerte Freigabe einer wirksamen Menge eines 
Arzneistoffs uber eine verlSngerte Zeitspanne, hergestellt in Mikrokapselform, worin die Zusammen- 
setzung umfasst: 

mindestens ein Polypeptid, bet dem es sich urn ein naturlich vorkommendes Luteinisierungs- 
hormon-freisetzendes Hormon (LH-RH), urn ein synthetisch hergestelltes Material vom gleichen Typ Oder 
urn synthetisch hergestellte Analoge von naturlich vorkommendem LH-RH handelt, die in gewisser Weise 
auf den Hypophysenvorderlappen unter Beeinflussung der Freisetzung von Luteinisierungshormon (LH) 
und follikelstimulierendem Hormon (FSH) wirken; 

ggf. mindestens ein Polymer-Hydrolysemodifizierungsmittel, ausgewahlt unter organischen Sauren, 
sauren Salzen, neutralen Salzen und basischen Salzen; und 

ein biokompatibles, bioabbaubares Verkapseiungspolymer, bei dem es sich urn ein Polylactid- 
Polymer, Polyacetal-Polymer, Polyorthoester-Polymer oder Polyorthocarbonat-Poiymer handelt. 

2. Zusammensetzung nach Anspruch 1, worin das Polypeptid ein Nonapeptid- oder ein Decapeptid- 
Analoges von LH-RH mit der Formel 

<pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z 

und den pharmazeutisch vertraglichen Salzen davon ist, worin: 

V Tryptophyl, Phenylalanyl oder 3-<1-Naphthyl)-L-alanyl ist; 

W Tyrosyl, Phenylalanyl Oder 3-{1-Pentafluorphenyl)-L-alanyl ist; 
X ein D-Aminosaurerest 

O 

— NH—CH— C— 
CH 2 
R 

ist, worin 

(a) ein carbocyclischer Aryl-enthaltender Rest, ausgewahlt unter Naphthyl, Anthryl, Fluorenyl, 
Phenylanthryl, Biphenylyi, Benzhydryl und Phenyl, substituiert mit drei oder mehr geradkettigen 
C,-4-Alkyigruppen, oder 

(b) ein gesattigter carbocyclischer Rest, ausgewahlt unter Cyclohexyl, substituiert mit drei oder mehr 
geradkettigen CWAIkylgruppen, Perhydronaphthyl, Perhydrobiphenyiyl, Perhydro-2,2-diphenylmethy1 
und Adamantyl, ist; 

Y Leucyl, Isoleucyl, Norleucyl oder N-Methyl-leucyl ist; 

Z Glycinamid oder — NH — R-, ist, worin C^-Alkyl, Ca- 6 -Cycloalkyl, Fluor-C,- 4 -alkyl oder 



12 



0 052 510 



o 

II 

— NH — C — NH — R 2 

5 ist, 

R 2 Wasserstoff Oder d.rAlkyt ist. 

3. Zusammensetzung nach Anspruch 2 mit einem Polymer, das ein Poly(lactid-co-glycolid)-Copolymer 
ist, worin das Copolymer Lactid-Glycolid in einem Molverhaltnis zwischen 100:0 und 40:60 umfasst und 
worin das Copolymer ein durchschnittliches Molekulargewicht zwischen etwa 20 000 und 100 000 hat. 
w 4. Zusammensetzung nach Anspruch 3, worin das Polypeptid in einer Menge zwischen 0,01 und 40,0 
Gewichtsprozent des Polymers vorhanden ist und das Hydrolysemodifizierungsmittel in einer Menge 
zwischen 1 und 15 Gewichtsprozent des Polymers vorhanden ist. 

5. Zusammensetzung nach Anspruch 3 oder Anspruch 4 mit einem Polypeptid, worin: 

V Tryptophyl oder Phenylalanyi ist; 
is W Tyrosyl ist; 

X 3-(2-Naphthyl)-D-alanyl oder 3-(2,4,6-Trimethylphenyl)-D-alanyl ist; 

Y Leucyl oder N-Methyl-leucyl ist; und 
Z Glycinamid oder NHEt ist; 

das Hydrolysemodifizierungsmittel Citronensaure, Ammoniumchlorid, Natriumchlorid oder Natrium- 
20 carbonat ist; und 

das Polymer Lactid-co-glycolid in einem Molverhaltnis zwischen 75:25 und 50:50 umfasst. 

6. Zusammensetzung nach Anspruch 5, worin das Polypeptid in einer Menge von 0,1 bis 10,0 
Gewichtsprozent vorhanden ist; 

das Hydrolysemodifizierungsmittel in einer Menge von 5 bis 10 Gewichtsprozent vorhanden ist; und 
25 das Polymer Lactid-co-glycolid in einem Molverhaltnis von 50:50 umfasst. 

7. Zusammensetzung nach einem der Anspruche 1 bis 6, worin das Polypeptid 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-0-alanyl-Leu-Arg-Pro-Gly-NH 2 

30 oder ein pharmazeutisch vertragliches Sauresalz davon ist. 

8. Zusammensetzung nach Anspruch 1, worin das Polypeptid ein Analoges von naturlichem LH-RH ist, 
wobei die Modification den Rest in der 6-Stellung betrifft, der von Gly in eine D-Aminosaure ubergefuhrt 
worden ist. 

9. Zusammensetzung nach Anspruch 8, worin die D-Aminosaure D-Ala, D-Leu, D-Phe oder D-Trp ist. 
35 10. Zusammensetzung nach Anspruch 9, worin die D-Aminosaure D-Leu ist. 

11. Zusammensetzung nach Anspruch 9, worin die D-Aminosaure D-Trp ist. 

12. Zusammensetzung nach einem der Anspruche 8 bis 11, worin die 10-Stellung modifiziert ist, so 
dass sich ein Nonapeptid in Form eines Alkyl-, Cycloalkyl- oder f luoralkylamins ergibt. 

13. Zusammensetzung nach einem der Anspruche 8 bis 11, worin das Gly-NH 2 durch ein 
40 Q-Azaglycinamid ersetzt ist. 

14. Zusammensetzung nach einem der Anspruche 8 bis 11, worin das Leucin in der 7-SteIlung durch 
N-Methyl-leucin substituiert ist. 

15. Zusammensetzung nach einem der Anspruche 7 bis 14, worin das Polymere ein Poly(lactid-co- 
glycol id )-Copoly meres ist. 

45 16. Zusammensetzung nach Anspruch 15, worin das Copolymere Lactid-Glycolid in einem Molver- 
haltnis zwischen 100:0 und 40:60 umfasst. 

17. Zusammensetzung nach einem der Anspruche 7 bis 16, worin das Polypeptid in einer Menge 
zwischen 0,01 und 40,0 Gewichtsprozent des Polymeren vorhanden ist. 

18. Zusammensetzung nach einem der Anspruche 7 bis 17, worin das Hydrolysemodifizierungsmittel 
so in einer Menge zwischen 0,1 und 20 Gewichtsprozent vorhanden ist. 

19. Zusammensetzung nach einem der Anspruche 7 bis 18, worin das Polypeptid in einer Menge von 
0,1 bis 10,0 Gewichtsprozent vorhanden ist. 

20. Zusammensetzung nach einem der Anspruche 7 bis 19, worin das Polymere Lactid-Glycolid in 
einem Molverhaltnis von 75:25 bis 40:60 umfasst. _ 

55 21 . Zusammensetzung nach einem der vorstehenden Anspruche in Form von injizierbaren Teilchen mit 
einer Grosse von etwa 1 bis 500 urn. 

22. Zusammensetzung nach einem der Anspruche 1 bis 21, die in einem pharmazeutisch vertraglichen 
Trager, der sich fur die parenterale Verabreichung eignet, dispergiert ist. 

23. Verfahren zur Herstellung einer Zusammensetzung nach einem der vorstehenden Anspruche, 
60 umfassend: 

Dispergieren einer wassrigen Losung, die das Polypeptid und ggf. ein Polymer-Hydrolyse- 
modifizierungsmittel in einem halogenierten organischen Losungsmittel enthalt, welches das Verkapsel- 
ungspolymer enthalt; 

Versetzen der Dispersion mit einem Koazervierungsmittel; und 
65 Sammeln der Mikrokapseln aus der Losung. 

13 



0 052 510 

Patentanspruche fur den Vertragsstaat: AT 



1. Verfahren zur Hersteliung einer pharmazeutischen Zusammensetzung fur die verzogerte Freigabe 
einer wirksamen Menge eines Arzneistoffs uber eine verlangerte Zeitspanne, hergesteilt in Mikrokapsel- 

5 form, dadurch gekennzeichnet, dass das Verfahren folgendes umfasst: 

Dispergieren einer wassrigen Losung, die ein Polypeptid, bei dem es sich um ein naturlich 
vorkommendes Luteinisierungshormon-freisetzendes Hormon (LH-RH). um ein synthetisch hergestelltes 
Material vom gleichen Typ Oder um synthetisch hergesteilte Analoge von naturlich vorkommendem LH-RH 
handelt, die in gewisser Weise auf den Hypophysenvorderlappen unter Beeinflussung der Freisetzung von 
w Luteinisierungshormon (LH) und follikeistimulierendem Hormon (FSH) wirken, und ggf. ein 
Polymer-Hydrolysemodifizierungsmittel, bei dem es sich um eine organische Saure, ein saures Salz, 
neutrales Salz Oder basisches Salz handelt, in einem halogenierten organischen Losung smittel, das ein 
biokompatibles, bioabbaubares Verkapselungspolymer, bei dem es sich um ein Polyactid-Polymer, 
Polyacetal-Polymer, Polyorthoester-Poiymer oder Polyorthocarbonat-Polymer handelt; 
is Versetzen der Dispersion mit einem Koazervierungsmittel und 
Sammein der Mikrokapseln aus der Losung. 

2. Verfahren nach Anspruch 1, worin das Polypeptid ein Nonapeptid- oder Decapeptid-Analoges von 
LH-RH mit der Formel 

20 (pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z 

und den pharmazeutisch vertraglichen Salzen davon ist, worin: 

V Tryptophyl, Phenylalanyl oder 3-(1-Naphthyl)-L-alanyl ist; 

W Tyrosyl, Phenylalanyl oder 3-(1-Pentafluorophenyl)-L-alanyl ist; 
25 X ein D-Aminosaurerest 

0 

— NH — CH — C — 

30 

CH 2 
R 

35 ist, worin R 

<a) ein carbocyclischer Aryl-enthaltender Rest, ausgewahlt unter Naphthyl, Anthryl, Fluorenyl, 
Phenylanthryl, Biphenyiyl, Benzhydryl und Phenyl, substituiert mit drei oder mehr geradkettigen 
C,-4-Alkylgruppen, oder 

(b) ein gesattigter carbocyclischer Rest ausgewShlt unter Cyclohexyl, substituiert mit drei oder mehr 
40 geradkettigen d- 4 -Alkylgruppen, Perhydronaphthyl, Perhydrobiphenylyl, Perhydro-2,2-diphenylmethyl 
und Adamantyl, ist; 

Y Leucyl, Isoleucyl, Norieucyl oder N-Methyl-leucyl ist; 

Z Glycinamid oder — NH— R, ist, worin R, C^-Alkyl, C 3 - 6 -Cycloalkyl, Fluor-CT^alkyl oder 
45 O 

— N H — C — N H — R 2 

ist, 

so R 2 Wasserstoff oder C^-Alkyl ist. 

3. Verfahren nach Anspruch 2, unter Verwendung eines Polymeren, das ein 
Poly{Iactid-co-glycolid)-Copolymeres ist, worin das Copolymere Lactid-Glycolid in einem Molverhaltnis 
zwischen 100:0 und 40:60 umfasst, und worin das Copolymere ein durchschnittliches Molekulargewicht 
zwischen etwa 20 000 und 100 000 hat. 

55 4. Verfahren nach Anspruch 3, worin das Polypeptid in einer Menge zwischen 0,01 und 40,0 
Gewichtsprozent des Polymeren vorhanden ist und das Hydrolysemodifizierungsmittel in einer Menge 
zwischen 1 und 15 Gewichtsprozent des Polymeren vorhanden ist. 

5. Verfahren nach Anspruch 3 oder 4 unter Verwendung eines Polypeptids, worin: 

V Tryptophyi oder Phenylalanyl ist; 
60 W Tyrosyl ist; 

X 3-{2-Naphthyl)-D-alanyl oder 3-(2,4,6-Trimethylphenyl)-D-alanyl ist; 

Y Leucyl oder N-MethyMeucyl ist; und 
Z Glycinamid oder NHEt ist; 

das Hydrolysemodifizierungsmittel Citronensaure, Ammoniumchlorid, Natriumchlorid oder Netrium- 
65 carbonat ist; und 



14 



0 052 510 



das Polymere Lactid-co-glycolid in einem Molverhaltnis zwischen 75:25 und 50:50 umfasst. 

6. Verfahren nach Anspruch 5, worin das Polypeptid in einer Menge von 0,1 bis 10,0 Gewichtsprozent 
vorhanden ist; 

das Hydroiysemodifizierungsmittei in einer Menge von 5 bis 10 Gewichtsprozent vorhanden ist; und 
5 das Polymere Lactid-co-glycolid in einem Molverhaltnis von 50:50 umfasst. 

7. Verfahren nach einem der Anspruche 1 bis 6, worin das Polypeptid 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphthyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 

io oder ein pharmazeutisch vertragliches Sauresalz davon ist. 

8. Verfahren nach Anspruch 1, worin das Polypeptid ein Analoges von naturlichem LH-RH ist, wobei die 
Modifikation den Rest in der 6-Stellung betrifft, der von Gly in eine D-Aminosaure flbergefuhrt worden ist. 

9. Verfahren nach Anspruch 8, worin die D-Aminosaure D-Ala, D-Leu, D-Phe oder D-Trp ist. 

10. Verfahren nach Anspruch 9, worin die D-Aminosaure D-Leu ist. 
is 11. Verfahren nach Anspruch 9, worin die D-Aminosaure D-Trp ist. 

12. Verfahren nach einem der Anspruche 8 bis 11, worin die 10-Stellung unter Bildung eines 
Nonapeptids in Form eines Alkyh Cycloalkyl- oder Fluoralkylamins modifiziert ist. 

13. Verfahren nach einem der Anspruche 8 bis 1 1, worin das Gly-NH 2 durch ein a-Azaglycinamid ersetzt 

ist. 

20 14. Verfahren nach einem der Anspruche 8 bis 11, worin das Leucin in der 7-Stel lung durch 
N-Methyl-leucin substituiert ist. 

15. Verfahren nach einem der Anspruche 7 bis 14, worin das Polymere ein Poly(lactid-co-glycolid)- 
Copolymeres ist. 

16. Verfahren nach Anspruch 15, worin das Copolymere Lactid-Glycolid in einem Molverhaltnis 
25 zwischen 100:0 und 40:60 umfasst. 

17. Verfahren nach einem der Anspruche 7 bis 16, worin das Polypeptid in einer Menge zwischen 0,01 
und 40,0 Gewichtsprozent des Polymeren vorhanden ist. 

18. Verfahren nach einem der Anspruche 7 bis 17, worin das Hydroiysemodifizierungsmittei in einer 
Menge zwischen 0,1 und 20 Gewichtsprozent vorhanden ist. 

30 19. Verfahren nach einem der vorstehenden Anspruche 7 bis 18, worin das Polypeptid in einer Menge 
von 0,1 bis 10,0 Gewichtsprozent vorhanden ist. 

20. Verfahren nach einem der Anspruche 7 bis 19, worin das Polymere Lactid-Glycolid in einem 
Molverhaltnis von 75:25 bis 40:60 umfasst 

21. Verfahren nach einem der vorstehenden Anspruche, worin das Produkt in Form von injizierbaren 
35 Teilchen mit einem Grossenbereich von etwa 1 bis 500 pm vorliegt 

22. Verfahren nach einem der Anspruche 1 bis 21, worin die Produktzusammensetzung in einem 
pharmazeutisch vertraglichen Trager dispergiert ist, der sich fur die parenteral Verabreichung eignet. 

Revendications pour les Etats Contractants: BE, CH, DE, FR, GB, IT, LI, LU, NL, SE 

40 

1. Composition pharmaceutique concue pour la liberation soutenue d'une quantity efficace de 
medicament pendant une pgriode prolonged, pr^paree sous forme de microcapsules, qui comprend: 

au moins un polypeptide qui est une hormone liberant I'hormone lutein isante (LH-RH) d'origine 
naturelle, une matiere du meme type preparee par synthese ou des analogues prepares par synthese de 
45 I'hormone LH-RH d'origine naturelle, qui agissent d'une certaine maniere sur la glande hypophysaire 
anterieure pour effectuer la liberation de I'hormone luteinisante (LH) et de I'hormone folliculo-stimulante 
(FSH); 

a titre facultatif, au moins un agent modificateur d'hydrolyse de polymere choisi entre des acides 
organiques, des sels d'acides, des sels neutres et des sels basiques; et 
so un polymere decapsulation biocompatible et biodegradable qui est un polymere du type polylactide, 
un polymere du type polyacetal, un polymere du type polyortho-ester ou un polymere du type 
potyorthocarbonate. 

2. Composition suivant la revendication 1, dans laquelle ledit polypeptide est un analogue 
nonapeptidique ou decapeptidique de I'hormone LH-RH, r§pondant a la formule 

55 

(pyro)Glu-His-V-Ser-W-X-Y-Arg-Pro-Z 

et ses sels pharmaceutiquement acceptables, formule dans laquelle 

V est un groupe tryptophyle, phenylalanyle ou 3-(1-naphtyl)-L-alanyle; 
60 w est un groupe tyrosyle, phenylalanyle ou 3-(1-pentafluoroph6nyl)-L-alanyle; 
X est un residu de D-amino-acide de formule 



65 



15 



0 052 510 



to 



25 



0 

— NH — CH — C — 
CH 2 
R 

dans laquelle R represente 

{a} un radical carbocyclique contenanl un groupe aryle, choisi entre ies radicaux naphtyle, anthryle, 
fiuorenyle, phenylanthryle, biph^nylyle, benzhydryle et phenyle substitu^ avec trois ou plus de trois 
groupes alkyle a chaine droite en d a C 4 ; ou 

(b) un radical carbocyclique sature choisi entre un radical cyclohexyle substitue avec trois ou plus de 
trois groupes alkyle en C, a C 4 a chaine droite, perhydronaphtyle, perhydrobiphenylyle, perhydro-2,2- 
; 5 diphenylmethyle et adamantyle; 

Y est un groupe leucyle, isoleucyle, norleucyle ou N-methyl-leucyle; 
Z est un groupe glycinamide ou un groupe — NH— R 1f dans lequel 

Rt est un groupe alkyle en d a C 4 , cycioalkyle en C 3 a C 6 , fluoralkyle en d a C 4 ou 

20 O 

— NH — C — NH — R 2 
R 2 est I'hydrogene ou un groupe alkyle en d a C 4 . 

3. Composition suivant la revendication 2, contenant un polymere qui est un copolymere du type 
poly(lactide-co-glycolide), ce copolymere comprenant des motifs lactide-glycolide dans un rapport molaire 
compris entre 100:0 et 40:60; et ce copolymere ayant un poids moleculaire moyen compris entre environ 
20 000 et 100 000. 

4. Composition suivant la revendication 3, dans laquelle ledit polypeptide est present en une quantity 
3° comprise entre 0,01 et 40,0% en poids du polymere; et ledit agent modificateur d'hydrolyse est present en 

une quantite comprise entre 1 et 15% en poids du polymere. 

5. Composition suivant la revendication 3 ou la revendication 4, contenant un polypeptide dans lequel: 

Y est un groupe tryptophyle ou phenylalanyle; 
W est un groupe tyrosyle; 

& x est un groupe 3-(2-naphtyl)-D-alanyle ou 3-(2,4,6-trimethylphenyl)-D-alanyle; 

Y est un groupe leucyle ou N-methyl-leucyle; et 
2 est un groupe glycinamide ou NHEt; 

ledit agent modificateur d'hydrolyse est I'acide citrique, le chlorure d'ammonium, le chlorure de 
sodium ou le carbonate de sodium; et 
40 ledit polymere comprend Ies motifs lactide-co-glycolide dans un rapport molaire compris entre 76:25 
et 50:50. 

6. Composition suivant la revendication 5, dans laquelle ledit polypeptide est present en une quantite 
de 0,1 a 10,0% en poids; 

ledit agent modificateur d'hydrolyse est present en une quantite de 5 a 10% en poids; et 
4 & ledit polymere comprend fes motifs lactide-co-glycolide dans un rapport molaire de 50:50. 

7. Composition suivant Tune quelconque des revendications 1 a 6, dans laquelle ledit polypeptide est le 
(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphtyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 ou un sel pharmaceutiquement 
acceptable de ce compost. 

8. Composition suivant la revendication 1, dans laquelle le polypeptide est un analogue de I'hormone 
so LH-RH naturelle, dans lequel la modification porte sur le changement du residu Gly en position 6 en un 

D-amino-acide. 

9. Composition suivant la revendication 8, dans laquelle le D-amino-acide est D-Ala, D-Leu,D-Phe ou 
D-Trp. 

10. Composition suivant la revendication 9, dans laquelle le D-amino-acide est D-Leu. 
55 11. Composition suivant la revendication 9, dans laquelle le D-amino-acide est D-Trp. 

12. Composition suivant Tune quelconque des revendications 8 a 11, dans laquelle la position 10 est 
modifiee en alkylamine, cycloaikylamine ou fluoralkyiamine pour offrir un nonapeptide. 

13. Composition suivant Tune quelconque des revendications 8 a 11, dans laquelle le Gly-NH 2 est 
remplace par un a-azaglycine-amide. 

60 14. Composition suivant I'une quelconque des revendications 8 a 11, dans laquelle la N-methyl-leucine 
remplace la leucine en position 7. 

15. Composition suivant Tune quelconque des revendications 7 a 14, dans laquelle le polymere est un 
copolymere du type poly(lactide-co-glycolide). 

16. Composition suivant la revendication 15, dans laquelle le copolymere comprend Ies motifs 
55 lactide-glycolide dans un rapport molaire compris entre 100:0 et 40:60. 



16 



0 052 510 

17. Composition suivant i'une quelconque des revendications 7 a 16, dans laquelle-ledit polypeptide 
est present en une quantite comprise entre 0,01 et 40,0% en poids du polymere. 

18. Composition suivant Tune quelconque des revendications 7 a 17, dans laqueile ledit agent 
modificateur d'hydrolyse est present en une quantite. comprise entre 0,1 et 20% en poids. 

s 19. Composition suivant I'une quelconque des revendications 7 a 18, dans laqueile ledit polypeptide 
est present une quantite" de 0,1 a 10,0% en poids. 

20. Composition suivant I'une quelconque des revendications 7 a 19, dans laqueile ledit polymere 
comprend les motifs lactide-glycolide dans un rapport molaire de 75:25 a 40:60. 

21. Composition suivant Tune quelconque des revendications precedentes, sous la forme de particules 
io injectables dont le diametre va d'environ 1 a 500 urn. 

22. Composition suivant Tune quelconque des revendications 1 a 21, qui est en dispersion dans un 
vehicule pharmaceutiquement acceptable propre a ('administration parenteral. 

23. Proc6d6 de preparation d'une composition suivant I'une quelconque des revendications 
precedentes, qui consists: 

'£ a dipserser une solution aqueuse contenant le polypeptide, et a titre facultatif, un agent modificateur 
d'hydrolyse de polymere, dans un solvant organique haiogene contenant ledit polymere d'encapsutation; 
a ajouter a ia dispersion un agent de coacervation; et 
a recueilir les microcapsules de cette solution. 

20 Revendications pour I'Etat Contra ctant: AT 

1. Procede de preparation d'une composition pharmaceutique sous forme de microcapsules, concue 
pour la liberation soutenue d'une quantite efficace de medicament pendant une periode prolongee, 
caracterise en ce qu'il consists : 

25 a disperser une solution aqueuse contenant un polypeptide qui est une hormone liberant I'hormone 
luteinisante (LH-RH) d'origine naturelle, une matiere du meme type preparee par synthese ou des 
analogues prepares par synthese de I'hormone LH-RH d'origine naturelle, qui agissent d'une certaine 
maniere sur la glande hypophysaire anterieure pour effectuer la liberation de I'hormone luteinisante (LH) et 
de I'hormone folliculostimulante (FSH), et a titre facultatif, un agent modificateur d'hydrolyse de polymere 

30 qui est un acide organique, un sel d'acide, un sel neutre ou un sel basique, dans un solvant organique 
haiogene contenant un polymere d'encapsulation biocompatible et biodegradable qui est un polymere du 
type polyiactide, un polymere du type polyacetal, un polymere du type polyortho-ester ou un polymere du 
type polyorthocarbonate; 

a ajouter a la dispersion un agent de coacervation; et 

35 a recueillir les microcapsules de cette solution. 

2. Procede suivant la revendication 1, dans lequel ledit polypeptide est un analogue nonapeptidique ou 
decapeptidique de I'hormone LH-RH repondant a la formule 

(pyro)Glu-His-V-Ser^/V-X-Y-Arg-Pro-Z - 

40 

et ses sels pharmaceutiquement acceptables, formule dans laqueile: 

V est un groupe tryptophyle, phenylalanyie ou 3-(1-naphtyl)-L-alanyle; 

W est un groupe tyrosyle, phenylalanyie ou 3-(1-pentafluorophenyl|-L-alanyle; 

X est un residu de D-amino-acide de formule 

45 

O 

— NH — CH — C — 
50 CH 2 

R 



dans laqueile R represente 
55 (a) un radical carbocyclique contenant un groupe aryle, choisi entre les radicaux naphtyle, anthryle, 
fluorenyle, phenylanthryle, biphenylyle, benzhydryle et phenyle substitue avec trois ou plus de trois 
groupes alkyle a chafne droite en C, a C 4 ; ou 

(b) un radical carbocyclique sat u re choisi entre un radical cyclohexyle substitue avec trots ou plus de. 
trois groupes alkyle en C, a C 4 a chaine droite, perhydronaphtyle, perhydrobiph^nylyle, perhydro-2,2- 
60 diphenylmethyle et adamantyle; 

Y est un groupe leucyle, isoleucyle, norleucyle ou N-methyl-leucyle; 

Z est un groupe glycinamide ou un groupe — NH — R 1t dans lequel 

R, est un groupe alkyle en G, a C 4 , cycloalkyle en C 3 a C 6 , fluoralkyle en C, a C 4 ou 



17 



0 052 510 



o 

— N H — C — N H — R 2 

5 R 2 est I'hydrogene ou un groupe alkyle en C t a C„. 

3. Proced6 suivant la revendication 2, utilisant un polymere qui est un copolymere du type 
poly(lactide-co-giycolide) qui comprend les motifs lactide-glycolide dans un rapport molaire compris entre 
100:0 et 40:60; le copolymere ayant un poids moleculaire moyen compris entre environ 20 000 et 100 000. 

4. Precede suivant la revendication 3, dans lequei ledit polypeptide est present en une quantite 
io comprise entre 0,01 et 40,0% en poids du polymere; et ledit agent modificateur d'hydrolyse est present en 

une quantite comprise entre 1 et 15% en poids du polymere. 

5. Procede suivant la revendication 3 ou la revendication 4, utilisant un polypeptide dans lequei: 

V est un groupe tryptophyle ou phenylalanyle; 
W est un groupe tyrosyle; 

is X est un groupe 3-{2-naphtyl)-D-alanyle ou 3-<2,4,6-trim6thylph6nyi)-D-alanyIe; 

Y est un groupe leucyle ou N-methyl-leucyle; et 
Z est un groupe glycinamide ou NHEt; 

ledit agent modificateur d'hydrolyse est I'acide citrique, le chlorure d'ammonium, le chlorure de 
sodium ou le carbonate de sodium; et 
20 ledit polymere comprend les motifs lactide-co-glycolide dans un rapport molaire compris entre 75:25 
et 50:50. 

6. Procede suivant la revendication 5, dans lequei ledit polypeptide est present en une quantite de 0,1 a 
10,0% en poids; 

ledit agent modificateur d'hydrolyse est present en une quantite de 5 a 10% en poids; et 
2$ ledit polymere comprend les motifs lactide-co-glycolide dans un rapport molaire de 50:50. 

7. Procede suivant Tune quelconque des revendications 1 a 6, dans lequei ledit polypeptide est le 

(pyro)Glu-His-Trp-Ser-Tyr-3-(2-naphtyl)-D-alanyl-Leu-Arg-Pro-Gly-NH 2 

30 ou un sel d'acide pharmaceutiquement acceptable de ce compose. 

8. Procede suivant la revendication 1, dans lequei le polypeptide est un analogue de I'hormone LH-RH 
naturelle, dans lequei la modification comprend le changement du residu Gly en position 6 en un 
D-aminoacide. 

9. Procede suivant la revendication 8, dans lequei le D-amino-acide est D-Ala, D-Leu, D-Phe ou D-Trp. 
35 10. Procede suivant la revendication 9, dans lequei le D-amino-acide est D-Leu. 

11. Procede suivant la revendication 9, dans lequei le D-amino-acide est D-Trp. 

12. Procede suivant Tune quelconque des revendications 8 a 11, dans lequei la position 10 est modifiee 
en line alkylamine, cycloalkylamine ou fluoralkylamine pour offrir un nonapeptide. 

13. Procede suivant Tune quelconque des revendications 8 a 11, dans lequei le Gly-NH 2 est remplac6 
40 par un a-azaglycine-amide. 

14. Procede suivant Tune quelconque des revendications 8 a 11, dans lequei la N-m6thyl-leucine 
remplace la leucine en position 7. 

15. Procede suivant I'une quelconque des revendications 7 a 14/ dans lequei le polymere est un 
copolymere du type poiy< lactide-co-glycolide). 

45 16. Procede suivant la revendication 15, dans lequei le copolymere comprend les motifs lactide- 
glycolide dans un rapport molaire compris entre 100:0 et 40:60. 

17. Procede suivant I'une quelconque des revendications 7 a 16, dans lequei ledit polypeptide est 
present en une quantite comprise entre 0,01 et 40,0% en poids du polymere. 

18. Procede suivant I'une quelconque des revendications 7 a 17, dans lequei ledit agent modificateur 
so d'hydrolyse est present en une quantite comprise entre 0,1 et 20% en poids. 

19. Procede suivant I'une quelconque des revendications 7 a 18, dans lequei ledit polypeptide est 
present en une quantite de 0,1 a 10,0% en poids. 

20. Procede suivant I'une quelconque des revendications 7 a 19, dans lequei ledit polymere comprend 
les motifs lactide-glycolide dans un rapport molaire de 75:25 a 40:60. 

55 21. Procede suivant I'une quelconque des revendications precedentes, dans lequei le produit est sous 
la forme de particules injectables dont les diametres sont compris dans un interval le d'environ 1 a 500 um. 

22. Procede suivant Tune quelconque des revendications 1 a 21, dans lequei la composition de produit . 
est dispersee dans un vehicule pharmaceutiquement acceptabje propre a ('administration parenterals 



65 



18 



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