JPH0338280B2 - - Google Patents

Description

[Detailed description of the invention]

The present invention provides a novel cyclosporin, a method for producing the same,
It relates to its use as a pharmaceutically active agent and to pharmaceutical compositions containing it. Cyclosporins are a group of structurally distinctive
It consists of a cyclic poly-N-methylated undecapeptide and has useful medicinal activities, especially immunosuppressive and anti-inflammatory activities. Cyclosporins include naturally occurring cyclosporins, such as cyclosporins A, B, C, D, and G, as well as chemically modified derivatives thereof, such as the corresponding dihydrocyclosporins (e.g., U.S. Pat. No. 4,117,118;
4210518 and 4108985 and German Patent Applications 2819094 and 2941080). Cyclosporins A, B, C, D and G are illustrative of that group and are conveniently represented by the following general formula. [In the formula, “MeC ₉ ” is a so-called “C ₉ -amino acid” residue represented by the general formula, (Here, the 2-, 3- and 4-positions are S
-, R- and R-coordination, and -X-Y-
is -CH=CH- (trans)), and A is -α-Abu- for cyclosporin A, -Ala- for cyclosporin B, and -Thr for cyclosporin C. -, in the case of cyclosporin D -Val-, and -NorVal in the case of cyclosporin G
− is. ] Throughout this specification, abbreviations such as -Ala
Amino acid residues represented by -, -Sar-, -MeVal-, -NorVal-, etc. are, according to conventional convention,
Unless otherwise specified, it should be understood that it has the (L)-coordination. Residue abbreviations beginning with "Me" represent N-methylated residues. The individual residues of the cyclosporin molecule are, according to the prior art,
Numbering is clockwise starting from residue 1-- _MeC9- . The same numbering system is used throughout this specification to indicate the position of individual residues. In the case of dihydrocyclosporin, −MeC ₉
-X-Y- of the residue (general formula) is -CH ₂ -CH ₂
− is. For example, in isocyclosporin, -
The MeC ₉ -residue is bonded to the -CO- group of the neighboring -MeVal- in the 11-position via its 0 atom in the 3-position instead of the N atom in the 2-position. Until recently, the methods of manufacturing cyclosporin were as follows:
Only fermentation methods and chemical modification of the resulting cyclosporin fermentation products were known.
However, fully synthetic methods are now available for the production of cyclosporine. This method is applicable to Japanese Patent Application No. 020779/1981 and European Patent Application No. 020779/1981.
No. 818100430 (publication number 0034567) and 1981
It is described in US Application No. 299103 filed on September 3, 2008, and the contents thereof are cited here in place of explanation. The basic feature of the total synthetic method is that, for the first time, (both cis and trans forms and dihydro form)
We have provided a means to obtain "C ₉ -amino acids", the corresponding [2R, 3S, 4S]-enantiomers, and N-desmethyl derivatives of these compounds in stereochemically pure form; It has not been previously known in an isolated or isolable state. According to total synthetic methods, these new compounds are used, usually in protected or activated form, as intermediates for the further synthesis of a series of open chain undecapeptides. Note that these undecapeptides should ultimately be cyclized to produce the product cyclosporin. Undecapeptide synthesis as well as cyclization is carried out according to the specific methods disclosed in the above references. Total synthetic methods therefore provide a means to obtain cyclosporins, enantiomers, etc. in which the " _C9 -amic acid" is substituted with the above-mentioned N-desmethyl derivatives. It has been found that, according to the invention, novel cyclosporins with useful pharmacological properties are obtained in which the residues in the 2- and/or 8-positions are replaced by amino acid residues of a defined type. The cyclosporin according to the invention is of the formula: [In the formula, Z ₁ is allyl or ethyl, Z ₂ is hydroxymethyl, or if Z ₁ is allyl, it is methyl, and -X-Y- is -CH=CH- (trans)
Or _-CH2 - _CH2- . ] A specific cyclosporin according to the present invention has the formula in which Z ₁ , Z ₂ and -XY- are as follows. () Z ₁ is allyl, Z ₂ is methyl and -X-Y-
is -CH=CH- (trans); () Z ₁ is ethyl, Z ₂ is hydroxymethyl and -X-Y- is -CH=CH- (trans); and () Z ₁ is ethyl, _Z2 is hydroxymethyl and -X-Y- is _-CH2 - _CH2- . Throughout this specification, these compounds will be referred to as ()
“Allylgly-cyclosporin”, () “(D)-Ser
−cyclosporine” and ()“(D)−Ser
-Dihydrocyclosporine" respectively. The cyclosporin according to the present invention can be produced by the total synthetic method described above. It was further surprisingly discovered that, with the above general formula -
Compounds in which X-Y- is -CH=CH- (trans) may also be used, for example, by the fermentation method disclosed for the production of cyclosporin A in the above-mentioned US Pat. It can be produced by adding amino acid precursors to the culture medium. And in this case, −Allylgly
- is in the cyclosporin molecule, specifically 2-
It was found that -(D)-Ser- is specifically incorporated at the 8-position. Cyclosporins according to the invention can be further subjected to chemical modifications or interconversions by methods known in the art. Therefore, from another point of view, the present invention also provides a method for producing cyclosporin according to the present invention, which method comprises the following steps. () (a) removing the O-atom protecting group from the cyclosporin according to the invention which is in O-atom protected form; or (b) removing the O-atom protecting group from the cyclosporin according to the invention and having the necessary sequence to obtain the cyclosporin according to the invention and as the N-terminus. The linear undecapeptide starting from the 8-position residue of cyclosporin (which undecapeptide is in unprotected or O-atom protected form) is cyclized and then, if necessary, (a) or () where -X-Y- is - as defined above.
For the production of cyclosporine in which CH=CH- (trans), (a) (L)- or (D, L)-allylglycine, (b) (D)- or (D, L)-serine, or ( c) cultivating a strain producing cyclosporin A in contact with a nutrient medium containing a mixture of amino acids (a) and (b) and then isolating the cyclosporin of the formula thus obtained from the culture; or () Cyclosporin according to the invention, for example (
The cyclosporin obtained according to a), (b) or () can be chemically modified, for example, when -XY- in the formula is -CH=CH- (trans) and Z ₁ is Hydrogenation of cyclosporine in which Z ₂ is hydroxymethyl, with the meaning given by -X-Y- in the formula -CH _2
-CH2- and _Z1 is ethyl to obtain the corresponding cyclosporin. Undecapeptides suitable for use in step (b) are e.g.
The above-mentioned Japanese, European and US patent applications
020779/1981, 818100430 and 299103 by combining a peptide sequence consisting of residues 8 to 11 of the cyclosporin molecule with a peptide sequence consisting of residues 1 to 7. However, in this case, the residues at the 2-position and/or the 8-position can be substituted with -Allylgly- or -(D)-Ser-, respectively, if desired. If a β-hydroxy-α-amino acid residue is present in the 8-position, it is suitably in the form with the O atom protected, for example in the form of an O-t-butyl derivative. The cyclization in step (b) is carried out by the special method described in the said patent application, in which the O atom protecting group is, for example, at the 8-position [step (
a)], this protecting group is finally removed according to methods known in the art of peptide chemistry. The general operations will be explained with particular reference to the synthesis of "(D)-Ser-cyclosporine" and its flowchart in Example 1 below. The preferred strain for use in the method of step () is Tolypocladium inflatum (G ams) sp.
The strain was NRRL 8044, and the culture was carried out by the United States Department of Penoria, Illinois, USA.
Agriculture (Northern Research and
Development Division), but is freely available to the public. Other cultures of this strain were given the code number FRI FERM-P No. 2796 and were consigned to the Microbial Technology Research Institute, Inage, Chiba City, Chiba Prefecture, Japan. The strain was originally Trichoderma polysporum (Link ex
Pers.) classified as belonging to a species;
Its morphological characteristics as well as the preparation and preservation of precultures and subcultures are fully described, for example, in US Pat. No. 1,491,509. According to the method of the present invention, a strain producing cyclosporin A was added (D)-
or (D,L)-serine, or added
In the presence of (L)-(D,L)-allylglucine or mixtures thereof,
1491509 or as described in Examples 2 and 3 below. The amino acid precursor is added at a concentration of about 1 to 1 per liter of culture medium.
It is suitable to add an amount of 15g, more preferably about 4 to about 10g, most preferably about 8g. Following culturing, the culture is harvested and the resulting cyclosporin of the formula, e.g. “(D)-Ser-
Alternatively, the "Allylgly-cyclosporin" is extracted according to known techniques, e.g. by comminution of the conidia and mycelia and then carrying out extractive and/or absorptive isolation. Separation of cyclosporine from other contaminants, especially contaminating cyclosporine A,
To achieve the separation from B, C, D and/or G, purification can be carried out, for example, by chromatography and/or recrystallization. Cyclosporin according to the invention, for example the step (
The cyclosporin obtained according to a), (b) or () is then, if necessary, subjected to chemical modification according to techniques known in the art, in particular by hydrogenation as in step (), e.g. In the case of “(D)-Ser-cyclosporin”, “(D)
If a β-vinylene-α-amino acid is present in the 2-position, it is simultaneously reduced to give -NorVal-, for example in the case of -Allylgly-. The hydrogenation is carried out in a manner analogous to known methods for reducing naturally occurring cyclosporins, such as cyclosporins C and D, to the corresponding dihydro derivatives, for example according to the general method disclosed in British Patent Specification No. 1567201. The hydrogenation can be carried out by catalytic hydrogenation at a neutral pH.
under conditions of temperatures of about 20 to about 30° C., pressures at or slightly above normal pressure, in the presence of an inert solvent or diluent, and in the presence of a catalyst such as platinum or preferably palladium (e.g., palladium on carbon). It is appropriate to carry out the Suitable solvents include ethyl acetate and lower fatty alcohols such as methanol and isopropanol. The resulting hydrogenation product can be purified by known methods, for example chromatographically. The following examples illustrate the method of the invention. Use the following abbreviations: BOC=t-butyloxycarbonyl Bzl=benzyloxycarbonyl-(D)-Ser(O-tBu)-=-(D)-Ser- in which the O atom is protected with t-butyl. -MeC ₉ = residue represented by the following formula. −MeDHC ₉ −=residue represented by the following formula. Example 1 Total synthesis of “(D)-Ser-cyclosporine” (1a) Boc-(D)-Ser(O-tBu)-MeLeu-
MeLeu−MeVal−MeC ₉ −Abu−Sar−MeLeu
-Val-MeLeu-Ala-Bzl. The title compound is prepared according to the flowchart shown on the following pages. Step A is carried out analogously to Examples 1m to 1r of the aforementioned Japanese, European and US patent applications with application numbers 020779/1981, 818100430 and 299103, respectively, and Step B is also carried out in Example 1a. It is carried out in exactly the same way as 1l to 1l. (1b) Boc−(D)−Ser(O−tBu)−MeLeu−
MeLeu−MeVal−MeC ₉ −Abu−Sar−MeLeu
-Val-MeLeu-Ala-OH. 3.56 g of the title compound of Example (1a) was dissolved in ethanol.
Dissolve in 70ml and add 13.2ml of 0.2N NaOH aqueous solution. The reaction mixture is left at 0° C. for 16 hours. Add 1N hydrochloric acid to the resulting clear liquid to adjust the pH to 5.
Evaporate to dryness under reduced pressure at 40°C. The residue is taken up in 300 ml of methylene chloride and 150 ml of demineralized water and shaken. After separating the organic phase, the aqueous phase is shaken with a further 150 ml of methylene chloride. The combined methylene chloride extracts are dried over Na ₂ SO ₄ , filtered through talc and the solvent is evaporated under reduced pressure. The residual product is purified by chromatography using 200 g of silica gel (0.062-0.2 mm) using methylene chloride/15% methanol as the eluent, and the eluted liquid is collected in 30 ml fractions. Distillate the title compound from 13 to 45
recovered in pure form from [α] ²⁰ _D = -164.7°, c
= 0.95 (in _CHCl3 ).

[Table] (1c) H−(D)−Ser(O−tBu)−MeLeu−MeLeu
−MeVal−MeC ₉ −Abu−Sar−MeLeu−Val
-MeLeu-Ala-OH. 1.9 g of the powdered product of Example (1b) are placed in 7 ml of trifluoroacetic acid precooled to -20 DEG C. and dissolved by swirling. The solution is left at -20 DEG C. for 11/4 hours and poured through a separatory funnel into a mixture consisting of ice, 200 ml of saturated sodium bicarbonate solution and 300 ml of methylene chloride. The aqueous phase must be non-acidic after shaking. After extraction twice with 300ml of methylene chloride,
The combined organic phases are dried over Na ₂ SO ₄ , filtered and evaporated. The residue was purified on silica gel (0.063-0.2 mm) using methylene chloride/15% methanol as the eluent.
Purify by chromatography on a 150 g column and collect the eluted solution in 25 ml fractions. The title compound is recovered in pure form from fractions 25-90. [α] ²⁰ _D =
−156°, c=0.96 (in _CHCl3 ). (1d) “(D)-Ser(O-tBu)-cyclosporin” 250 mg of the product of Example (1c) was added to 10 methylene chloride in a 50 ml flask fitted with a calcium chloride tube.
Dissolve in ml. Add 70.4 mg (0.077 ml) of N-methylmorpholin dissolved in 1 ml of CH ₂ Cl ₂ and then
Add 49.4 mg of propanefluoric anhydride dissolved in 1 ml of CH ₂ Cl ₂ . The reaction mixture is stirred at 22°C for 21 hours. Dilute the resulting solution with 200 ml of methylene chloride and add 100 ml of sodium bicarbonate solution in a separatory funnel.
Shake with ml. Methylene chloride aqueous solution again
Extract with 100 ml, dry the organic phase over Na ₂ SO ₄ , filter and evaporate the solvent. The residue is purified by chromatography on 150 g of silica gel (0.063-0.2 mm) using methylene chloride/5% methanol as eluent and the eluted liquid is collected in 25 ml fractions. The title compound is recovered in pure form from fractions 13-14. [α] ²⁰ _D = -203°, c = 0.95 (in CHCl ₃ )
. (1e) “(D)-Ser-cyclosporin” 345 mg of the product of Example (1d) was added to 2 ml of trifluoroacetic acid.
Dissolve in and leave at 0°C for 6 hours. The reaction mixture was diluted with 150 ml of methylene chloride and placed in a separatory funnel on ice/
Shake with 100 ml of bicarbonate of soda solution. Separate the alkaline raw aqueous phase and re-extract with 200 ml of methylene chloride. Combine the methylene chloride extracts with Na ₂ SO ₄
Dry on top, filter and evaporate the solvent completely under vacuum. The residue was purified with silica gel (0.063-0.2 mm) using methylene chloride/5% methanol as the eluent.
Purify by chromatography on top of g, and the eluted solution is
Collect fractions of 25ml each. Distillate the title compound
Recovered in pure form from 42 to 52. R _f [chloroform/acetone (1:1)] = 0.39. Example 2 Total synthesis of “(D)-Ser-dihydrocyclosporin” The title compound was converted into −MeDHC ₉ in place of −MeC ₉ − at the 1-position in the flowchart of Example (1a).
It is manufactured in the same manner as in Example 1 except that - is used. The products of the last three steps of the synthesis have the following properties. (a) Boc−(D)−Ser(O−tBu)−MeLeu−MeLeu
−MeVal−MeDHC ₉ −Abu−Sar−MeLeu−
Val−MeLeu−Ala−OH: [α] ²⁰ _D = −153°, c
= 0.85 (in chloroform). (b) H−(D)−Ser(O−tBu)−MeLeu−MeLeu−
MeVal−MeDHC ₉ −Abu−Sar−MeLeu−Val
−MeLeu−Ala−OH: [α] ²⁰ _D = −154.8°, c=
0.92 (in MeOH). (c) “(D)−Ser(O−tBu) −
dihydrocyclosporin”: [α] ²⁰ _D = −205°, c =
1.03 (in _CHCl3 ). (d) “(D)−Ser −dihydrocyclosporin”: [α] ²⁰ _D
= −241°, c = 1.0 (in _CHCl3 ). Example 3 Microbiological production of (D)-Ser-diciclosporin (3a) Per liter, 50 g of glucose, 8 g of (D)-serine, _0.75 g _{of K2HDO4} , _{MgSO4.7H2O0.5}
10 liters of nutrient medium containing g, 0.1 g of CaCl ₂ .6H ₂ O and 8 g of ammonium malate are inoculated with 1 liter of the conidial and mycelial suspension of strain NRRL 8044 obtained from the preculture. Inoculate medium into 100 Erlenmeyer flasks.
ml at a time, then at 27°C for 14 days at rotational speed.
Incubate with a stirrer at 180 rpm. The mycelium is separated from the culture medium and extracted using a Turrax apparatus by grinding and stirring twice with 4.5 liters of 90% methanol. The ground mycelium is separated from the solvent by suction and treated again twice with 90% methanol as described above. Combine the liquids and heat to 40℃ under vacuum.
The vapor is evaporated and concentrated until it consists mainly of water. The resulting mixture is saturated with common salt and extracted three times with 3 liters of ethyl acetate each time, then the ethyl acetate solutions are combined, dried over sodium sulfate and concentrated by evaporation under vacuum at a temperature of 40°C. . The resulting residue is chromatographed on 440 g of silica gel (Merck silica gel, particle size 0.04-0.063 mm) using chloroform/2% methanol as eluent and collected in 300 ml fractions. Fractions 41 to 53 were combined and evaporated under vacuum,
The residue (242 mg) was then dissolved in 1 ml of methanol, and purified using a high-pressure chromatography column (RP8, diameter 8 mm, length 25
cm, temperature 70℃, flow rate 10ml/min, pressure 160 atm)
The mixture was chromatographed in 125μ aliquots to obtain 77mg of pure “(D)-Ser-cyclosporin”.
R _f [chloroform/acetone (1:1)]=
0.39. NMR spectroscopy confirms the identity of the product of Example (1e). (3b) Preculture bacteria for the above method are produced as follows. The suspension of spores and mycelium used for inoculation was prepared from a culture of the initially isolated strain NRRL 8044, containing 20 g of malt extract per liter of demineralized water;
Culture on an agar medium containing 20 g of agar and 4 g of yeast extract at 27°C for 21 days. The spores of this cultured bacteria were taken into physiological saline solution and the final concentration was 5×10 ⁶
spores/ml. 10ml of this suspension
, with the exception of the (D)-serine component, was used to inoculate 1 liter of a nutrient solution with the same composition as the culture medium of Example (3a), and the culture was incubated at 27°C for 3 days on a rotary shaker (200 rpm). It will be carried out. Use the fermentation solution as inoculation medium for the final culture. Example 4 "Allylgly - Microbiological production of cyclosporin" 50 g glucose per liter, 8 g (D,L)-allylglycine, 0.75 _{g K2HPO4} , _MgSO4 .
The strain was added to a nutrient medium containing 0.5 g of 7H ₂ O, 0.1 g of CaCl ₂ 6H ₂ O and 8 g of ammonium malate.
NRRL 8044 (manufactured by the method described in Example 3)
Inoculate 500 ml of pre-cultured bacteria. Fill 50 Erlenmeyer flasks with 100 ml each of the inoculating medium.
It is then cultured at 27° C. for 14 days with a stirrer at a rotation speed of 180 rpm. Isolate mycelium from filter culture,
Using a Turrax apparatus, extract by grinding and stirring twice with 4.5 liters of 90% methanol. The ground mycelium is separated from the solvent by suction and treated again twice with 90% methanol as described above. The liquids are combined and evaporated under vacuum at a temperature of 40°C until the vapor consists mainly of water. The resulting mixture is saturated with common salt and extracted three times with 3 liters of ethyl acetate each time, then the ethyl acetate solutions are combined, dried over sodium sulfate and evaporated under vacuum at a temperature of 40°C. The resulting residue is chromatographed on 320 g of silica gel (Merck silica gel, particle size 0.04-0.063 mm) with chloroform/2% methanol as eluent and fractions of 0.5-1.0 liters are collected. Fractions 9 and 10 are combined and evaporated under vacuum to give 2.9 g of an oily residue. The residue was chromatographed on 240 g of silica gel using chloroform/1-2% methanol as eluent and collected in 20 ml fractions. Fractions 257 to 261 were combined and evaporated under vacuum to give semi-pure " A residue consisting of 460 mg of "Allylgly-cyclosporine" is obtained. This residue was dissolved in 1 ml of methanol, water (35%)/methanol (15%)/acetonitrile (50%) was used as the eluent, and a high-pressure chromatography column (RP8, diameter 8 mm, length 25 cm, temperature 70°C, A flow rate of 10 ml/min and a pressure of 160 atm) were passed through 125 microns each to obtain 191 mg of concentrated "Allylgly-Cyclosporin". The last chromatographic step is repeated to obtain 87 mg of "Allylgly-cyclosporin" in pure form. R _f [Silica gel on glass, chloroform/acetone (1:1)] = 0.69 [Silica gel on glass, chloroform/methanol (9:1)] = 0.66 Cyclosporin according to the invention can be used as shown in the following standard test: , exhibiting useful pharmacological, especially immunosuppressive and anti-inflammatory activities. 1 Immunosuppressive activity 1.1 Local hemolysis phenomenon in vitro in gel [RIMishell
and RWDutton, J. Exp. Medicine, 126,
423-442 (1967)]. Cyclosporine according to the invention has a concentration of 0.1
to 10.0 μg/ml shows a hemolytic inhibition zone compared to untreated controls. 1.2 Lymphocyte stimulation test according to Janossy and Greaves [Clin.Exp.Immunol., 9, 483, (1971) and 10, 525 (1972)]: Cyclosporine according to the invention is administered at a concentration of 0.3
Concanavalin A-stimulated RNA synthesis (H ³ -
(inhibition of thymidine assimilation), cell proliferation and embryonic development in mouse splenic lymphocytes. 2. Anti-inflammatory activity Anti-inflammatory activity can be demonstrated in the Freund's supplement arthritis latency test in rats and the Freund's supplement arthritis-treatment test in rats when administered in appropriate amounts. In view of its immunosuppressive activity, cyclosporine according to the invention is desirable for use in the prevention and treatment of diseases and conditions requiring a reduction in the immune response. Accordingly, the cyclosporins of the invention can be used to suppress the proliferation of lymphocytes and immune cells, for example in the treatment of autoimmune diseases or in preventing rejection of transplants, such as skin, bone marrow and kidney transplants. In view of its anti-inflammatory activity, cyclosporine according to the invention is suitable for use in inflammatory conditions such as arthritis and polyarthritis chronica.
It is desirable to use it in the treatment of rheumatic diseases such as progrediente. The toxicity level of the novel cyclosporins according to the invention is generally about the same or slightly less than that of the known commercially available cyclosporins obtained from natural sources. In accordance with the foregoing, the present invention provides, in another aspect, a cyclosporin of the formula as defined above for use as a medicine, i.e. for therapeutic use, and in particular for use in therapeutic or prophylactic methods as defined above. do. For the aforementioned uses, especially as an immunosuppressant, the required daily dosage is in the range of about 50 to 900 mg, such as about 75 to 600 mg, preferably about 200 to 400 mg, and suitable for oral administration. The dosage form consists of about 25 to 300 mg of cyclosporin mixed with a solid or liquid pharmaceutical diluent or carrier. Preferred cyclosporins according to the invention, namely “Allylgly-cyclosporin” and “(D)-Ser
- Cyclosporine” in 1.1 to 1.3 above.
The results obtained from the test are shown below.

In accordance with the foregoing, the present invention also provides a pharmaceutical composition comprising a cyclosporin as described above, particularly a cyclosporin of the formula, and a pharmaceutically acceptable diluent or carrier. Preferred pharmaceutical compositions according to the invention can be made using cyclosporin according to the invention as a monocyclic peptide according to known methods, for example according to the technique of JP-A-54-132223. The outline is as follows. Preferred formulations according to the invention contain, for example, pharmacologically active cyclosporine as active ingredient and the following ingredients: (a) transesterification products of natural vegetable oil triglycerides and polyalkylene polyols, (b) ethanol and (c) vegetable oils. A pharmaceutical composition comprising a carrier. Component (a) can be prepared by conventional methods, for example as disclosed in US Pat. No. 3,288,824. The ester may be the transesterification product of two mole parts of a natural oil such as corn oil, almond oil, peanut oil, olive oil and/or palm oil and one mole part of polyethylene glycol having a molecular weight of 200-800. Such esters are available under the trade name LABRAFIL (Fieder, Lexicon der Hilfstadtsfe) from France, Boulogne sur Seine, Etabrisman Gattohotse.
Hilfstoffe, p. 320, 1971]. Preferred esters are those derived from unsaturated natural oils, such as Labrafil, which is a triglyceride oleate polyethylene mixture.
It is M1944CS. (Labrafil is a trademark.) The compositions described above may also include one or more other ingredients, such as (d) saturated fatty acid triglycerides. Such ingredient (d) can be prepared in a conventional manner. In general, these glycerides have an iodine value of less than 2. Examples of such glycerides include Bitten/Ruhr, commercially available under the trade name MIGLYOL, especially MIGLYOL 812 from Deinamit Nobel, or under the trade name MIRITOL 318 from Henkel, Düsseldorf, West Germany. (Miglyol and Militol are trademarks.) The preferred total concentration of the components present in the pharmaceutical composition according to the invention as well as the weight ratios of the individual components will depend inter alia on the individual components used, especially their solvent/solubilizing effect, the individual cyclosporine,
It will depend on the concentration of cyclosporine desired in the final composition and the solvent/solubilizing effects of other pharmaceutical excipients present. Generally preferred component (a) [and
If present, the weight ratio of (d) to peptide is equal to all components.
10 parts to 0.2 to 10 parts of peptide, or preferably 1 to 10 parts of peptide, preferably 1 part of peptide by weight.
~7 parts by weight. If components (a) and (d) are both present, component (a)
The weight ratio of component (b) to component (b) may be, for example, from 1:1 to 1:2. The pharmaceutical composition of the present invention comprises pharmacologically active cyclosporin as previously defined components (a), (b) and (c).
It can be produced by mixing with a liquid carrier consisting of [and other optional components such as (d)]. If one of the ingredients is a solid, temperatures up to about 70° C. may be used to produce a liquid melt in which the active ingredient can be dissolved. The composition can be cooled and then, for example, ground. The pharmaceutical composition of the present invention can be formulated into a form suitable for oral or parenteral administration by a conventional method, optionally with pharmaceutical excipients. Preferably they are in liquid form. A preferred composition is as follows. (a) Drink solution, such as Example 1, (b) Drink emulsion, (c) Injection solution, (d) Solution filled into capsules. Components (b) and (c) may be present in a combined concentration of up to 60% of the total composition. The content of ethanol as component (b) is, for example, 2 to 5% for parenteral compositions and 1 to 5% for oral compositions, calculated based on the total composition.
It may be 20%. The content of component (c), vegetable oil, such as olive oil or corn oil, in both oral and parenteral compositions is, for example, 35-60% by weight relative to the total weight of the composition. The production of a drink solution as an example of a formulation is as follows. Mixture of Labrafil M1944CS and ethanol (weight 40:
15) Dissolve in 1 ml at 25℃. Add 0.4ml of olive oil or corn oil. Filter the resulting mixture;
Fill into small vials. The final solution contains 10 parts by weight of Labrafil, 3 parts by weight of cyclosporin, 3 parts by weight of ethanol and 5 parts by weight of olive oil or corn oil.

Claims (1)

[Claims] 1 Cyclosporin represented by the formula. [In the formula, Z ₁ is allyl or ethyl, Z ₂ is hydroxymethyl, or if Z ₁ is allyl, it is methyl, and -X-Y- is -CH=CH- (trans)
Or _-CH2 - _CH2- . 2. Cyclosporin according to claim 1, which is "(D)-Ser-cyclosporin". 3. Cyclosporin according to claim 1, which is "(D)-Ser-dihydrocyclosporin". 4. Cyclosporin according to claim 1, which is "Allylgly-cyclosporin". 5. An immunosuppressant containing cyclosporine represented by the formula as an active ingredient. [In the formula, Z ₁ is allyl or ethyl, Z ₂ is hydroxymethyl, or if Z ₁ is allyl, it is methyl, and -X-Y- is -CH=CH- (trans)
Or _-CH2 - _CH2- . ]