CH640521A5 - Cyclosporin D - Google Patents

Abstract

Cyclosporin D of the formula I and possessing an antiarthritic effect. <IMAGE>

Description

The present invention relates to cyclosporin D of the formula I.

CH3v ^ ^ CHj CH

I CHj ^ yCH3

CH2 CHj CH

'' -L

L t.

CH3 \

c II

\ h2

Ir

HO ^ R ^ CH ^ CH

9h3 1

CH3n / CH3

CH3 CH

ch3-n-ch-co-n

C ti-

II o

CH3

CH CO N CH c (Si CH-

CH-

ch3

\

<

/

co

Ch-ch2-ch l

H

I! O

CO

n-ch.

CHj- N

H

I °

OC-CH—

I.

CH3

• N CO-CH—

i I

H CH3

-CO CH-

CH2 CH-i

I.

CH / \ CH3 CH3

O

II

L

c-

—Ch-1

1

ch

H

h n-co ch

CH-j CH3

CH 2

I.

CH3 CHj and pharmaceutical preparations.

Cyclosporin D is obtained by producing a cyclosporin D-producing strain of the fungal species Tolypo-

3rd

640521

cultivates cladium inflatum chamois in the presence of a nutrient medium and isolates cyclosporin D.

The breeding can be carried out according to methods known per se for the breeding of analog strains.

A preferred strain producing cyclosporin D is the freely accessible strain NRRL 8044 of the fungal species Tolypocladium inflatum Gams. A culture of these was deposited in the United States Department of Agriculture (Northern Research and Development Division), Peoria, III., USA. This strain was previously assigned to the mushroom species Trichoderma polysporum (Link ex. Pers.) And is e.g. described in DOS 2 455 859.

Strains of the fungal species Tolypocladium inflatum Gams can also be used for the production of cyclosporin D, e.g. by selection or mutation of the fungal strain NRRL 8044 under the influence of ultraviolet or X-rays or by application of other measures, e.g. by treating laboratory cultures with suitable chemicals.

Cyclosporin D can be isolated in a manner known per se. Cyclosporin D can be separated from natural products that are present in larger quantities, e.g. the more polar cyclosporin A (also known as S 7481 / F-1), the more polar cyclosporin B (also known as S 7481 / F-2) and the even more polar cyclosporin C.

Cyclosporin D is characterized by interesting chemotherapeutic and pharmacological properties and can therefore be used as a remedy.

Cyclosporin D is particularly indicated as antiarthriti-cum.

Cyclosporin D can be administered as a remedy alone or in a suitable pharmaceutical form with pharmacologically indifferent auxiliaries.

In the following example, all temperatures are given in degrees Celsius.

Example cyclosporin D.

500 liters of a nutrient solution containing 40 g glucose, 2.0 g sodium caseinate, 2.5 g ammonium phosphate, 5 g per liter

MgS04. Containing 7H20, 2 g KH2P04, 3 g NaNOs, 0.5 g KCl, 0.01 g FeS04 and demineralized water are inoculated with 50 liters of a preculture of strain NRRL 8044 and in a steel fermenter with stirring (170 5 rpm) and aeration (1 liter air / min. / Liter nutrient solution) incubated at 27 ° for 13 days (see DOS 2 455 859).

The culture broth is stirred with the same amount of n-butyl acetate, after the organic phase has been separated off, it is concentrated in vacuo and the crude extract is degreased by 3-stage distribution between methanol-water (9: 1) and petroleum ether. The methanolic phase is separated off, concentrated in vacuo and the crude product is precipitated by adding water. The material obtained after the filtration is chromatographed on silica gel with hexane-Ace-15 ton (2: 1) as the eluent, the fractions first eluted predominantly containing cyclosporin A and cyclosporin D, the later eluted fractions predominantly being cyclosporin C. For further purification the fractions containing cyclosporin A and D are crystallized from the 2 to 20 2.5-fold amount of acetone at −15 ° and the crystals are then separated further by double chromatography on silica gel, the fractions cyclosporin D eluted first with water-saturated ethyl acetate in strong contain enriched form. These are dissolved in twice the amount of acetone and left to crystallize at -15 °. The crude crystallizate of cyclosporin D thus obtained is dissolved in 10 times the amount of acetone for further purification, 2% by weight of activated carbon is added and the mixture is heated to 60 ° for 5 minutes. The clear and almost colorless filtrate obtained after 30 filtration through talc is concentrated to a third of the volume and allowed to cool to room temperature, with cyclosporin D spontaneously crystallizing out. The crystallization is completed by standing at -17 °. The crystals obtained by filtering are washed with a little ice-cold acetone and then dried in a high vacuum at 80 ° C. for 2 hours.

Characterization of Cyclosporin D 40 Colorless, prismatic crystals; Mp 148-151 ° [a] D20 = -245 ° (c = 0.52 in chloroform)

[a] D20 = -211 ° (c = 0.51 in methanol).

v

Claims (3)

640521 2nd PATENT CLAIMS 1. Cyclosporin D of Formula I, CH3n ^ CH I. CHj I. C. Ü Ir l! Ov R / Cl-. " CR CH3 ^ CHj CH 3. / CH3 ch3-N-CH-CO-N- chn I I * ch c n L Ii o ch i ch3 ch I. ! -ch- c 0 — u - 1 -ck - c- l 1 L II H O CHa ch3 CH- co ch-ch2-ch t_ co w-chn CK3-N H OC-CH N CO-CH - N CO CH 1 I ch3 H I. CHj -N C i I CH 2 CH3 I. CH CH3 CH3 H I. : H - N — CO C ri CH CH3 <~ H3 i CMj! CH CHj CH3 2. Pharmaceutical preparations containing cyclosporin D. 35