JPH0446120A - Arteralization inhibiting agent - Google Patents

Abstract

PURPOSE:To provide a low toxic arteralization-inhibiting agent containing a herbimycin derivative as an active ingredient. CONSTITUTION:The objective agent contains a compound of formula I (R<1>, R<2> are Cl, or both are bonded to each other, when R<1>, R<2> are Cl, R<3> and R<4>, R<5> and R<6> both are bonded to each other, R<7>, R<8> are H; when R<1> and R<2> are bonded to each other, R<3> is OCH3, Cl, R<7> is H, group of formula II, R<3> is H, Br, R<4> is OCONH2, R<5> and R<6> form O or a bond together, or R<4> and R<5> form O-CO-O together and R<6> is Br) as an active ingredient. The compound is useful for preventing or treating various diseases caused by the abnormal multiplication of blood vessels, such as rheumatic arthritis, diabetic retinopathy, immature baby retinopathy, senile macular degeneration and excessive cicarrix formation when wound is healed.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to an angiogenesis inhibitor. Angiogenesis inhibitors are expected to be a preventive or therapeutic agent for diseases caused by abnormal proliferation of blood vessels, such as rheumatoid arthritis, diabetic retinopathy, retinopathy of prematurity, senile macular degeneration, and excessive scar formation during wound healing. be done.

Conventional technology Substances that inhibit angiogenesis include, for example, medloki/progesterone, sulfated polysaccharides, and crude bovine cartilage extract, and the combination of cortiscin and heparin can inhibit angiogenesis. It is also known that it can be done.

Herbimycin is an antibiotic classified as an ansafine antibiotic with herbicidal activity, anti-tobacco mosaic virus activity, P3880 Ikemia, B16 Melanoid 7.112
It is known to exhibit antitumor activity in mouse experimental animal systems using 10 leukemia, Lewis Lang carcinoma, and Ehrlich acytes carcinoma. Certain derivatives of herbimycin are known as Ehrlich γ.
Cytes carcinoid 7 is known to have antitumor activity in a mouse experimental animal system [Journal Antibiotics (J, Antibiotics)].
otics) 37.1264 (I984): 39
．． 415 (I986)].

It is known that herbimycin A and its derivatives differentiate cancerous cells into normal cells [Molecular and Cellular Biology (Mol,
Ce1l, Biol, ), 6, 2198 (I986
) Journal on Antibiotics (J.

Antib+ot+cs), 41.831 (I988
)].

Problems to be Solved by the Invention An object of the present invention is to provide a new angiogenesis inhibitor useful as a pharmaceutical.

Means for Solving the Problems According to the present invention, the general formula (I) (wherein R1 and R2 represent Cp or together represent a bond, and when R1 and R2 are Cf, R3 and R4o and R5 and R6 each together represent a bond,
R'ii and R8 represent H. When R' and R2 together represent a bond, R3 is 0CR3 or Cf
, R7 represents H or N, and R8 represents 14
or Br, and R'', R5 and R6 together represent 10- or a bond when R4 is DCON), R4 and R5 together represent -o-co- When R6 represents o-, R6 represents Br.

1'F shows specific examples of compound (I) and their respective physicochemical properties.

(I) Compound J-] TLCRf value = 051 (Benzene: Ethyl acetate 1゛1) Melting point 138t 7-spectrum m/Z 590 (M”, [ff0)1
4J2[110) Hydrogen nuclear magnetic resonance spectrum (CDCf
3 middle, middle) δ(92m)6, 98(l)I, dd
, J= ~1.0.11.5Hz), 6.59(I
H, ddJ=2.0.2. (Ez), 6.46 (IH
, dd, J=N, 5, 11.5 Hz).

5, 80 (Ift, dd, J=11.5.11.5
Hz), 4.56 (I, H, d J = 11.5 Hz > 4, 17 (IN, s).

2, 96 (Ill, d, J = 9.0Hz).

t) Compound I TLCRf value: 0.60 (benzene: ethyl acetate-1゛1)C)l, 011 Ultraviolet absorption spectrum: λ++axnm (ε) 24
5<12,000) Mass spectrum m/z 629 (
M-, C+Jt7NJs) Hydrogen nuclear magnetic resonance spectrum (
in CDCf3) δ (ppm) 7.60 (l) I,
brd), 6.48(IN, s>,
4.48 (IH, brs) Compound I TLCRf value: 0.63 (benzene acetone-7,3
) Melting point: 188℃ (decomposition) C1l, Oil Ultraviolet absorption spectrum: 2□8 mouth m (ε) 2
32 (I8,500) high resolution mass spectrum: m
/z 578.239([”,,113.[:βN20
Calculated value as 6 578.239) Hydrogen nuclear magnetic resonance spectrum ([: in DCA3) δ (ppm) 7.23 (
LH, d, J=2.3Hz), 6.60(IN,
dd.

J=2.3.3.0)tz), 5.89(IN, d
d, j=7.61.1.6Hz)5.80(LH,b
rs), 5.51 (LH, qd, J=1, 0.7.
1Hz)5.10(I)1. brd, J・7.6H
z), 4.50(I)1. d, ,b3.0Hz)
, 1.66 (3H, d, J=1.111z) Elemental analysis: C60,01, H6,92, N 4.71°Cz
9L, [l] Calculated value as N206: C60,
18,116,80,N 4.84° (4) Compound 1-
4 Cf 5゜89 ○ TLCRf value: 0,80 (Benzene diacetone 7:3
) Melting point: 199°C (decomposition) C8, Oil Ultraviolet absorption spectrum, λ. nm(ε) 2
71 (23,500) high resolution mass spectrum: m
/z 553.200 (calculated value as CaeLt[:I!2NO6 553.200) Hydrogen nuclear magnetic resonance spectrum (CDCA, medium): δ (ppm) 7.33 (IH
, d, J=2.5Hz), 6.63(IH, ddJ
=2.0.2.5Hz), 6.55(E, d, J
= 13.5Hz) 5.86 (IH, dd, J = 9.8
．． 13.5Hz), 4.99 (IH, dd.

J=27 10.6Hz), 4.66(IH, dd,
J=2.7.9.8Hz) 1, 75 (3)! ,
d, J=1.3Hz) Elemental analysis: C60,28,H
6,98,N 2.45. Cf 1289C281 (
,, (J, calculated value as NG6: C60°74.
H6,74,N 2.53. [:ffl 12.6
4(5) Compound ■-D TLCRf value: 0.45 (benzene: acetone-7,3) Melting point: 178°C (decomposition) Ultraviolet absorption spectrum: 8a+t nm (E) 258
(I8,600) Hydrogen nuclear magnetic resonance spectrum (CDCf
3) δ (ppm) 6.92 (E, d, J = 0.9
Hz), 6.42 (IH, qd.

J-11, 11, 5Hz), 6.32 (I8, dd
, J211.5°11.5flz), 5.30
(IN, dd, J=IO, 6, 11, 511z
) 5.28 (IH, qd, J=1.0. 9.8t
lz), 5.03 (IH, d.

J=9.4Hz), 4.49(IN dd,
J=0.9)1z), 4.00(I1(,dd,
J29.1. IQ, 6tlz>, 3
1B(it(,dd.

J-1,8,1O,0Hz), 2.25(I)1.
m), 1.26 (3H, dJ=1.0Hz) Elemental analysis: C54,89, H6,32, N 4.26
．． 8r 12.68C3oH+ Calculated value as 18rLOs: Mouth 55.20. H6,34,N 4.29.
8r 12.10 (6) Compound I-6 TLCRf value 0.84 (benzene, acetone = 7:3) Melting point: Optical rotation 132°C (decomposition) [αJ, = 13° (cO, 5, mouth H [β3) CH,O
ll Ultraviolet absorption spectrum: λma,, nm (ε) 26
8 (I8,000) hydrogen nuclear magnetic resonance spectrum (CDI
J3) δ (99m) 7.28 (IH, Qd,
J21.2. 12.2Hz), 6.81(IH
.

d, J=1.6Hz), 4.6H1)1. d,
J=7.3Hz), 4.47(IM, s), 4.
32 (LH, d, J=9.4Hz), 2.32 (I
H, m).

],, 83 (3H,s) Elemental analysis: C48,97,H5,31,N 1.93
．． Br 21.69[:3oLsCalculated value as BrNO+o: C49,24,H5,38,N 1.92.
Br 21.16 Compounds l-1 and I-2 are published in JP-A-63-218620, and compounds 1-3 to I-6 are published in Journal on Antibiotics (J, ^n
These are known substances described in tib+ot+csL39 4.15 (I, 986) together with their respective manufacturing methods.

Compound (I) can be administered in the form of tablets, granules, powders, capsules, syrups, ointments, creams, injections, etc. prepared by conventional methods, depending on the purpose and method of administration. It is particularly preferable to use it in the form of an injection. When used as an injection, the compound (I
) at 1 to 1000 g/kg per day.
Administer intravenously or locally in ~3 doses.

Compound (I) has angiogenic activity and is less toxic than no-1-bimycin A. In addition, since the compound does not have the same effect on existing blood vessels as herbimycin A, it is effective against various diseases caused by abnormal proliferation of blood vessels, such as rheumatoid arthritis, diabetic retinopathy, retinopathy of prematurity, and senile. It is useful for macular degeneration, excessive scar formation during wound healing, etc. Next, the toxicity and pharmacological effects of the compound (H) will be explained using test examples.

Test Example 1 Acute Toxicity Test A test compound suspended in physiological saline containing 2% gum arabic was administered intraperitoneally to 6-week-old male DDY mice (25 ± 1 g, 3 mice per group). 50 from the survival rate after hours
As a result of calculating the % survival dose (LD5゜) using the increasing/decreasing method, all compounds (I) had LDso: >200 mg/
It was kg.

Test Example 2 Inhibitory effect on angiogenesis in the pickled membrane of chicken fertilized eggs N.
Tanari et al.'s method [Experimental Bathology (E
Experimental Pathology) 30
．． 143 (I986) El, the effect of compound (I) on angiogenesis in the pickling membrane of fertilized chicken eggs was investigated.

A small hole is made in 10 to 20 4.5-day-old fertilized chicken eggs.
Vinyl acetate-ethylene copolymer (Eν40;
A test compound encapsulated in a tube (manufactured by Mitsui-Dupont) was installed,
After culturing in a machi egg container at 37°C for 2 days, add 10% sesame oil pore agent Lointralipos (+ntral+pos); Midori Jujisha! ! ] 1- was injected into the pickled membrane, and the extent of the inhibitory effect of the test compound on blood vessels generated within the pickled membrane was observed.

That is, eggs with 3 or more areas in which no blood vessel formation was observed were treated as eggs that showed complete inhibition, and the inhibition rate was calculated using the following formula.

Total number of chicken eggs (number of eggs) In addition, the p value was determined by Fischer's exact method. The p value for all data was 0.05 or less.

The results are shown in Table 1. As shown in Table 1, the compound of the present invention exhibited a significant angiogenesis inhibitory effect gE't+ when administered at 0.01 to 10 mg/unit.

Test Example 3 Effect experiment on existing blood vessels Using 10-day-old VI sinuses, a test compound was placed and cultured in the same manner as in Test Example 2, and the effect on blood vessels already formed within the pickled membrane was investigated. . Table 1 shows cases in which disappearance of existing blood vessels was observed and cases in which no change was observed.

No. table The present invention will be illustrated below with examples.

Example After dissolving 200 g of injection compound (L-1) in ethanol 201, it was sterilized by pressure filtration using a millibore filter (pore size 022 μ). The resulting sterile p-liquid5. Oml
was dispensed into brown vials, freeze-dried by a conventional method, and
Obtain mg/vial of lyophilizate.

Effects of the Invention The present invention provides a new angiogenesis inhibitor useful as a pharmaceutical.

Claims (1)

[Claims] General formula (I) ▲There are mathematical formulas, chemical formulas, tables, etc.▼ (In the formula, R^1 and R^2 represent Cl or together represent a bond, and R^1 and R^2 If is Cl, then R^
3 and R^4 and R^5 and R^6 each represent a bond together, and R^7 and R^8 represent H. R
If ^1 and R^2 together represent a bond, then R
^3 represents OCH_3 or Cl, R^7 represents H or ▲There are mathematical formulas, chemical formulas, tables, etc.▼, and R^8
represents H or Br, R^4, R^5 and R^6
is R^5 and R^6 if R^4 is OCONH_2
together represent -o- or a bond, and when R^4 and R^5 together represent -O-CO-O-, R^6 represents Br. ) An angiogenesis inhibitor containing a herbimycin derivative represented by the following as an active ingredient.