JPH0314581A - Antibiotic cc-1065 fragment - Google Patents

Abstract

NEW MATERIAL: Compounds of formula I [R₁ is H, CH₃, -CH₂Ph, CH₂=CH₂-, -CH₂SCH₃, -CH₂OCH₃, -CH₂OCH₂CH₂OCH₃, -CH₂CCl₃ or -CH₂CH₂Si(R₂)₃; R₂, R₂' and R₃ are each H, a 1-5C alkyl or phenyl; R₄ is SO₂R₂, SO₂CH₂CO phenyl or CO₂CH₂Z; X is OSO₂R₂, Cl, Br, I or OH].

EXAMPLE: 4,5-Pyrrolo-6-methoxyindoline.

USE: An antibacterial agent, bacteria controlling agent for washed clothes, bacteria controlling rinsing agent for impregnated paper and fabric, useful for controlling growth of sensitive bacteria in plate certification method and microorganism culture medium.

PROCESS: A compound of formula II is reacted with a reagent comprising an alkanesulfonyl chloride, carbon tetrachloride/triphenylphosphine, carbon tetrabromide/triphenylphosphine, and N-iodosuccinic imide/triphenylphosphine.

COPYRIGHT: (C)1991,JPO

Description

DETAILED DESCRIPTION OF THE INVENTION Antibiotic CC-1065 is disclosed in US Pat. 16
No. 9,888, disclosed and claimed by chemical and physical variables. Then antibiotic cc-1065
The structure of “Structure verification of antibiotic CC-1065 (S
StructureProof of Amtibiot
ic cc-1065) J DG March Y (D.G, Martin), C.G.
C.G. Chidester, D.J. Ducham
p), and S.A.Mizusaku (8, A.Miz
aak), J, Antibiot, Volume 33, 902
Page (1980), it was clarified.

The structure of antibiotic CC-1065 is shown in E1. The antibiotic CC-1065 consists of a system of three fragments, the most unstable part of the molecule being 1,2,8.85L-cyclop[C]penzo(1,2-1):4.3- 1)'] Dipyrrole-4 (5H) monoone, book E! In Book A, m this is shown as the compound tla. Antibiotic CC-10
Attempts to obtain this fragment failed due to the loss of yield of 65. For this reason, certain intermediates disclosed in the specification may be caused by certain bacteria, such as Bacillus saccharis.
ubtilis), Klebsiella pneumoniae, Sarcina-A/
Sarcina lutea, Staphylococcus aureus, and Mycobacterium avium (Myc
obacterium avium ) 'I. These compounds can therefore be used to disinfect washed and stacked tableware contaminated with Staphylococcus aureus. Additionally, the antimicrobially active compounds of the present invention can be used as a bacteriostatic rinse agent for laundered clothing, and for impregnated paper and fabrics, and to inhibit the growth of susceptible bacteria in plate assays and microbial cultures. is also useful. Generally, the antimicrobially active compounds of the present invention are described in U.S. Pat.
69. It can be used in the same manner as disclosed for antibiotic CG-1065 in No. 888. These uses are well known in the antibiotic art. This microbiological technique is therefore readily available to those skilled in the art for such applications.

As mentioned above, an 11-step method for making compound Cl2 is shown in page 2. This stage is the following and bp.

Step 1: First step of combination (nucleophilic substitution of aromatic)
J, Bou-raais
) and C, Mahieu, Comp
t, Redux (C), vol. 263, p. 84 (1966
year). Also, G. Bourdais and C. Germain, Te
See also t, Letters 195 (1970). Various R groups can be introduced onto the phenol precursor of (1) by procedures described in the literature (appropriate references detailed under Step 8). The various malonates, β-ketoesters and β-diketones used are all known compounds.

Stage 2 - Reduction. When R2=alkoxy, diisobutylaluminum hydride is the screening reagent. The reaction conditions are very specific for optimal yields (see Reference Example 1). When Rz=alkyl or phenyl, standard reduction procedures using sodium borohydride can be used.

Step 3 Monofunctional group exchange. In particular, the chemistry described herein is
This is the case when X = So2CH3. For example, mesylate or tosylate can be used with other acid acceptors such as pyridine (with and without the addition of a solvent such as methylene chloride), or trialkylamines (with a solvent) and the corresponding sulfonyl chloride. can be made under standard conditions known to the art. 40I-rogen analogs can be made by standard procedures known in the art, such as Ph3P/CCm(cnr4) and N-iodosuccinimide/tri7enylphosphine.

Stage 4: Unification of reduction. This step is a new method for the preparation of indolines (dihydroindoles). This involves reduction of the nitro to amino group and concomitant intramolecular cyclization, yielding (5). The reduction step described in detail herein involves H2 in alcohol in the presence of a tertiary amine;
I am using pto2.

These are the standard hydrogenation conditions for this technology. Palladium or nickel catalysts can also be used, and bases other than tertiary amines, such as pyridine, can also be used. In the reduction situation of Alternative 9, Fe or T i c l3 or Sn in the acid
Ct2 can be used. This would then require another step involving base treatment to cause cyclization to (5). An example of reduction by iron is Fe/CH, CO2H/
Use CH3CH20H (GS Ponticello)
G., S., Fonticello) and G.G. Baltouiy ( :f, J, Balaw
in),゛. T, Org, chem. Volume 44 4
Page 003 (1979)- ) a Rz = ca,
These conditions are required in the case of ph or -CH2CH=CE2.

The idea of nitro reduction followed by in situ cyclization to indole was developed by A. D. Batcho.
and Da Stage 5 W, Leim-gruber (W, Leim-gruber)
ber), German Patent Publication No. 2057840 (
The reductive cyclization to indole was advanced by Reissert (1971).
) procedure (R, J. Sundberg),
``Iy }'-The Chemistry
7 of engineering naoles) J pages 176-183,
(See Academic Press, New York, 1970).

Substitution of one unstable group. This step is necessary because X is incompatible in the chemical reaction of step 8. This converts X to acetate or C to C under standard conditions (alkali carboxylate in acetone, DMF or alcohol).
It consists of substituting the conjugate base of the alkino ν carboxylic acid of No. 5. Since some hydrolysis may also occur when x=So2CH3, the reaction mixture is treated with acetic anhydride before f61 alone.

Step 6 - Nitration with acetic acid, acetic anhydride, sulfuric acid, deoxyacid/
It can be carried out under various conditions described in the literature including nitric acid in H20, 7-alcohols and hinitroalkanes. The four-spectral selectivity of this reaction is supported by the spectroscopic data obtained.

The reduction of the i9i7-2l-portion to the amino group follows the description of the chemical reaction in step 4, except that the base is omitted.

Step 8 - Indole combination. This procedure is outlined in sumannindole chemistry [P.G.Gasman (P.G.Gasman)
ssman) et al., J. Am+Chsm, Soc,
g6 volume, pages 5494, 5508, 5512 (197
4 years)]. Gassmann's work requires several changes not disclosed or suggested. The chemical sequence and some of the intermediates are shown in Figure 3. α−
Thiomethyl esters are known. This method uses hachlorosulfonium complex A, which is combined with aniline (6
), it deviates from the published Gasmann route. Gassmann made chloroamine of anilineb and reacted it with thioether to form oxindole. Second, the method uses two different bases, while Gasmann uses two equivalents of aniline followed by two equivalents of base.
use. Triethylamine, diisopropylethylamine, bis(1.8L) is preferably bis(1,8-
di/thylamine) + "phthalin, and the base 2 is triethylamine. Chloroform, acetonitrile,
A variety of solvents can be used, such as tetrahydrofuran (THIF) and methylene chloride, the latter being preferred.

The temperature range is from 1000 to -800, and the reaction is carried out under an inert atmosphere. The cyclization to oxindole B is carried out by acid catalysis ( 2 N H
Ct, Step 9 ether and/or ethyl acetate).

The final reduction (reductive removal) to (9) can be achieved with lithium aluminum hydride (as described by Gassmann and $-1 or diborane type reagents, with the latter being overwhelmingly superior.

A preferred reagent is (CH
3) 28-BH3.

- This deprotection v1 stage # (removal of R1) is Rl in Example 8.
= Described in detail in CH3.

Many procedures have been described in this art, including the cleavage of methyl ether, but under an inert atmosphere (95-100°)
hexamethylphosphoric triamide (EMPA)
) was found to be effective only among the alkylmercazotides (z
x ・C ・+7z Luchi (S, C, Welch) and A.S.C.P.Rao (A.S, C, P,
Rao), Tet, Letters No. 505 (1
977) and T.R.Kelly
lly), H, M, Dali (H, M, Da
li) and W.G. Tour 7 (W.G.
．． Tliang), Tet. Letters, 335
No. 9 (1977), or Me2 in dichloroethane
S-BBr3 (P.G. Willard)
．． Willard) and CB-7 Lyle (C
．． B, Fryhle), Tet, Letters
No. 3731 (1980)].

When R,=C112Ph, standard hydrogenolysis conditions (H2, Pd/C) are well suited for deprotection (
Org, Reactions Volume 7, Page 263 (1
953)]. When Rl=CH,S CH3, mercuric chloride in acetonitrile/H20 removes the ether (R.A. Holton)
ton) and R.G. Tevis (R.G.D.
avis), Tet. Letters No. 533 (
(1977)]. ．． When R1: CH20CH3, a mild acid such as acetic acid produces phenol 10 (
J. Med. Chem. 9, p. 1 (1966) or Syntbegi 8 244 (1976)]. But in reality, this protecting group is lost in step 6, but under standard conditions prior to step 7? Can be introduced. R■=-an
When 2oca2ca20CH, phenol is ca
ZnBr2 or TiC in 2cz2 (T
et, Letters No. 809 (1976)].

When Rl=-CE2ca=CH2, several two-step procedures carry out the deprotection of the ether [Pi in alcohol
/C, Ang, Chem, Engineering nt. Ed, Volume 15 5
p. 58 (1976); Sea2, CH in dioxane
3C (72H, Tet, Letters No. 2885 (1970); t-BuOK, DMSO followed by H2S04 in acetone, , r, Chem, So
c, 1g03 (1965): RhC in alcohol
t(PPh3)z, DABCO followed by I) H2, . T
, Org. Chem, vol. 38, p. 3224 (1973)
Year)〕. In the case of R1a -CH2CH2S i (R2)3, deprotection is carried out with Bu, NF [H, Gerlach et al., Hslv. C
hin, Acta. Volume 60, page 3039 (1977)
].

See step 101 #t3.

Step 11 - This step (when X=Bro) is facilitated by contacting with silica gel and! It also occurs when left in a protic solvent. Is this reaction the first? The process also proceeds in the presence of an aqueous solution of a base such as a class amine, pyridine, t-poxide, etc., or a weak base such as a carbonate or a charcoal salt.

The following examples are illustrative of the products and methods of the invention, but are not to be considered limiting. Unless otherwise noted, all percentages are by weight and all proportions of solvent mixtures are by volume.

亙Δ2巳 Aryl diethyl malonate (2) to 2-
Preparation of aryl-1,3-propanediol (3) Diisoptylaluminium hydride (M
BAL) 100y (0.70 mol) added to the name. To this stirred solution, add arylmalonate f21 in THF IQ■ 33. Add Uf (0.105 mol). The addition rate is adjusted to keep the reaction temperature below 5°.

After the addition is complete, remove the water bath. After a total reaction time of 3 hours, the reaction is stopped by adding the solution in portions to cold 3 N HCt (approximately 1.5 1 ) with stirring.

? Add 1 t of EtOAC to the mixture followed by "H2Ct2
Extract 1000 meters. The combined organic phase was treated with Na2
Dry over S04 and concentrate to a reddish-brown residue (21.1)t. Residual silica dal 500? 60% EtO on top
Chromatography with an eluent gradient of Ac/hexane → 100% KtOAc revealed that the diol +31 (I
T-62.598) n. 7y (49% yield) as a pale red oil (hardens on standing in cold K).

NMR (CDCz3): 7.5 ~7. o (
m, 3H), 3.80 (s, 3H), 4.
0-3.3 (including m. 7H-201'!) MS
, C1oH13NO5 Calculated value: 227.07
94 Measured value: 227.0780 Analysis and calculated value: C, 52.86, H, 5.76
．． N, 6.16 Measured value: C, 53.40. H
, 5.77. N. 57.99■2-Alli Lou L
2-aryl 1,3 from 3-propanediol (3)
-Dero/ξandiolpismenre} (4) Preparation N
O diol in 100 d of dry pyridine (
Add methanesulfonyl chloride 6.89 (0.06 mol) to 34 4.7 F (0.2 mol) with stirring. Add at 5", divide (a) at room temperature, and then cool.
The solution was concentrated in vacuo and CH2Ct2/iN HC4
It will be featured inside. The organic phase is separated, dried over Na2So, and concentrated to the residue. Trituration with EtOAc yields an off-white solid, and the mother liquor is purified by silica gel (
Chromatography on KtOAc eluent)
Total yield 16.655' (yield 86), melting point 122-12
3'' (recrystallized from acetone) to yield compound (4) bismesylate (U-62,597).

NMR (Acet-d6): 7.7-7.2
(m. 3H), 4.62 (d.4H,.
T=JHz), 4.11 (t.IH, J=7Hz),
3.92 (8, 3H), 3.06 (S.5
H).

Sei Tetsu: Calculated value for CL2H 7NOg82 C.
37.59, H, 4.47, N, 3.65 measured value C, 37.35, H, 4.44, N. 3
．． 59 This compound was assayed against L 1210 Mus musculus leukemia cells in culture in a standard in vitro dilution assay with the following results.

Dso (μg/+d)=6. o ID9σ (μg/
rnt) = 18 Volume Example 3 Preparation of 2-aryl-1,3-propaneshiol bismesylate (4) Kara 6-methoxyindoline bismesylate (5) THF 30 -
nt: 20 ml of EtOAc, and anhydrous:x:.
．． jl / -le 150 Compound in rILt (4i
Add triethylamine 1.5- and pt02400 to 1.915' (0.005 mol). While shaking this solution, the hydrogen pressure was increased to 0.492-0.703Ka.
/crIL2 (7-10 psi)
. The reaction solution is then filtered over Celite and concentrated in vacuo. After azeotroping with CH2Ct2 several times in vacuo, the residue is finally taken up in CH2 (:t2100 rnl and cooled under N2 in an ice bath. 1.5 ml of triethylamine is added to the stirred solution. Then, add 900 μt of methanesulfonyl chloride dropwise.After stirring, bring the solution to room temperature for ω minutes.Next, add the solution I N H
Wash with Ct, dry over Na2So, and concentrate. The residue was rapidly chromatographed on silica 1501 with an eluent of 60% KtOAc/hexane 500% followed by 80% eluent 1000% as an off-white solid, mp 122-123' (recrystallized from ethanol). ) compound (5), 6-methoxyindoline bismesylate (a
-s2,ss6)1.3y (yield 78%) was recovered.

DMF-dヮ” 7.36 (d+I
H, J=8.5[1z) ,7.00((L, l
H, J=2[IZ), 6.69 (d(1.1
H. J=2. 8.5Hz). 4.47 (2
H. (L.J=6Hz), 4.3-3.6 (
m, 3H). 3.80 (8.3H). 3.20
(11.3H), 3.07 (8,3H).

Analysis: CxzHx? For NSzoa, the calculated value C,
42.97, H. 5.11, N, 4.18 Measured Value C, 42.87. H. 5.27. N,
4.29 This compound was tested by a standard test tube dilution assay.
The culture medium was assayed against L1210/- murine leukemia cells, and the following results were obtained.

Engineering D. (μg/a/) = 4.8 engineering D9. (μg/
J=jO Emperor△1yu e-Methoxyi. indoline bismesylate (5) to 6-methoxyindoline acetate (
6) Preparation of 6-methoxyindoline bismesylate (5) in DMF 3Q 13. Of (39 mmol), absolute ethanol 800- followed by sodium acetate 321
Add.

The heterogeneous mixture is refluxed under N2 for 24 hours, cooled and concentrated in vacuo. The residue is treated with 100% acetic anhydride for 2 hours (with stirring at room temperature) and then concentrated in vacuo. The residue is taken up in CH2CL2/H20, the organic phase is separated, dried over Na2So, filtered through charcoal and concentrated to an oil residue. When the oil becomes solid, compound (6) 6
- yielding methoxyindoline acetate 11.62 (
If necessary, further purification is possible by silica glucomatography using 60% EtOAC/hexane eluent).

NMR (CDCt3): 7.17 (d,L
H, J=8.5[1z), 7.02(a, IH
, J=2Hz), 6.60 (eL6. IH,!
=2. 8.5+Iz), 4.18 (a,
2 H. J=6Hz). 4.1-3.4 (m.3H
), 3.78 (8.3EI). 2.91 (8,3H),
2.05 (S.3H).

Analysis For C engineering 3H engineering 7 No. S, calculated value: C, 52.16. Hl 5.76, M
．． 4.68 Measured value: c, 52.13, H, 5
．． 79, N, 5.27 thoughts: Calculated value: 299. υ
827 Measured value: 299.0823 Ema'' 6-Methoxyintolerine acetate (6)
Preparation of 5-nitro-6-methoxyindoline acetate (7) from 500η (1.67 mmol) of 6-methoxyindoline acetate (6) in 20 parts of nitromethane (90% H)
Add 90μt of NO. The cooled (0-5°) reaction solution is separated, dried over Na2So, and concentrated.

The residue was chromatographed on Nricadel 502 (ωφ
EtOAc/hexane eluent → 100% F2tOAc
) to give compound (7), 5-nitro-6, as a yellow solid, melting point 175-177' (recrystallized from ethanol).
-Methoxyintrine acetate (Tlr-62,69
6) Produces 440η (yield 76%).

Hentan (DMy-a.): 7.91 (8.1 H
), 7.20 (e. lH). 4.27 ( ”
r2H, J=6Hz), 4-3 3-7(J3
H). 3.98 (8. 3H), 3.17 (s.
3H). 2.07 (8.3H).

9j death: C13”16N2073 Calculated value for K
, 45.34, H, 4.68, N, 8.14
Measured value C, 44.81, H, 4.77, N,
8.16 Test this compound in a standard test tube dilution assay.
It was assayed against L 1210 murine leukemia cells in culture with the following results.

ID,. (μg/d)=>Cut D,. (μg/d)=
>Fi(1ffi± Preparation of 5-amino-6-methoxyindoline acetate (8) from 5-nitro-6-methoxyindoline acetate (7) 5-nitro-6-methoxy in THF 5Q and anhydrous ethanol 150 rnt Indoline acetate (7) 4.5P (13
Mi! J-T: Le) [Add Ptoz 500 1W,
”20.703KqlcrrL2 (10p81),
Shake at l (about ω minutes) until ingestion stops. Filter and concentrate in vacuo. After concentration, product 3.01 precipitates out. This was filtered off and the mother liquor was rapidly chromatographed on silica glue 1007 with KtOAe eluent, adding 0.6
2.

Total yield (3.6) of 5-amino-6-methoxyindoline acetate (U-62,697) of compound (8)
r) is 88%7, melting point 134-135', (may be acetone/cyclohexane).

NMR (CDCt3)' 7.02 (8,j
H), 6.65 (“+ I H), 4.16
(d. 2H, J = 6 [1z), 4.1 -3
．． 5 (m, 3H). 3.83 (S, 3H),
3.6(br.B,2H)+2. 83
(sr3H).

Analysis 20. H. Calculation is accurate for N20,S C. 49.67, H, 5.77, N,
8.91 measured value c, 49.74, H. 5.7
2. H, 8.94 This compound was assayed against L 1210 Mus musculus leukemia cells in culture by a standard in vitro dilution assay with the following results.

Engineering D. (μg/mz) =>Cut D9. (μg/+
++/) => Father Example i Preparation of 4,5-pyr6ro-6-methoxyindoline (9) from 5-amino-6-methoxyindoline acetate (8) Drying at -75 under nitrogen CH2Ct214III1! niC
l2/CH2Cl, solution (2r)ttL Cld m
l CE2Ctz ) 6.0 ml wo7JIll
t le. Add CH3(CH3S)CH to the stirred solution.
C02C2H5 370 μt (2.5 mmol) [E. H. Wick (LH. Wick), T. Yamaniθhi, H. C.
Wasai? (JC, Wertheimer)
, Y-E, Hoff, B.F. Proctor
and Q: S.A. Gold Prix (S, A, Go
ldb+lith), J, Agr. Food Che
m. According to the procedure in Vol. 9, p. 289 (1961), CH.
3(Br)CHC0202H, prepared from tomethylmercadetide]. After 5 minutes, dry CH2Ctz 3.
〇1,8-bisdimethylaminonaphthalene 47o
A solution of wq (2.2 mmol) toani17/indoline(8)628η (2.0 mmol) is added dropwise over 15 minutes. The red solution is stirred for 2 hours at -7 Da, then 350 .mu.t of triethylamine in 65 .mu.t of CHzCtz is added dropwise over a few minutes. Remove cooling bath.

When the reaction solution reaches room temperature, it is briefly concentrated in vacuo. Add 5 ml of EtOAC, Keiichitel 51, and 6 ml of 2 N HCL to the residue and stir vigorously for 2 hours. Separate the organic phase and dilute the aqueous layer with EitOAc/
Extract with Et,O (1:l). The organic phases are combined, dried over Na2So, and concentrated. At this point the residue was taken up in THF IQd and 'l M BH3.
Treat with SMe2 3 , Q ml under nitrogen overnight at room temperature. Alternatively, the diastereomeric oxidants derived from the acid treatment were at this point subjected to silica chromatography (50%, f'i0% E
itOAc/Hexane or 9Q% BtOAc
/hexane).

GC-MS: m/e M+414 (15%), 2
27 (100%)-2' 118E-30 NMR (CDCt3): 8.4 (+)r.
8, IH), 7.11 (8. II{), 4.
5-3.7 (m. 5 H), 3.90 (
8. 3H), 2.95(s, 3H). 2.10(a,
3H), 1.92 (sl 3H). 1.82 (8,3H
) - major diastereomer (2.5/1): minor diastereomer shows the following differences: -1.99 for 〇CH3 and 1CH3
(8. 3H), 1.76 (e. 3a), N
Hpa7.7. The CE2 band lies between 4.3 and 3.6 ppm.

The aqueous phase from acid treatment is neutralized and extracted with CH2Ct2.

The C H2C t2 solution is dried, concentrated, and chromatographed on silica group with 50% acetone/cyclohexane, resulting in a recovery of >40% starting material (anilinoindoline) and 2 fl deacylated starting material. .

The borane dimethyl sulfide reductive removal reaction (to B) is worked up by stopping with 1 N HCz at 1 when gas evolution ceases and taken up in CH,Ct2/H20.

The separated organic phase is dried over Na2So and concentrated.

Chromatography of the residue on silica gel (M% acetone/cyclohexane) yielded bioproduct (9) 15
5 1Ni is produced. Simple yield 75%, based on recovered starting material 85%. Melting point 182-18.3” (
Phase change at 160', recrystallization from chloroform). Product 4,5-pyrrolo-6-methoxyinthophosphorus (9) (
U-62, 233).

NMR (CDCt3): 8.3 (br,
s, IH), 6.96 (B. 2H). 4.
2-3-5 (m.5 H+OH) l3. 9
2 (s, 3H). 2.87 (81 311:),
2.41 (8.3H).

Analysis For C4HIEIN204S, calculated value C,
54.17, H, 5.84, N, 9.03 measured value C, 53.49, H, 5.96, N,
9.42GC-MS: O-acetate m/e M
+352 (13%). 213 (1υO article)-2'-
1 reward SE-addition, temperature 150-260°C lo 7 minutes),
single peak.

The compound was assayed against L 1210 Mus musculus leukemia cells in culture by a standard in vitro dilution assay;
I got the following results.

Dsa (μg/ml!) =>4 D90 (μ
g/+y)=)4 Example 2 4. Preparation of 4.5-pyrrolo-6-hydroxyindoline (10) from s-pyrrolo-6-methoxyindoline (9) To 10rj of dried and degassed HMPA (hexamethyl m-acetic acid triamide) was added 35ml of butylmercabutane at room temperature.
Add 0 μt. Cool the solution in an ice-water bath and add 1,5 M n-BuLi 2,Otd in hexane dropwise. The reaction was cooled down to room temperature 1 and then introduced (9).
Add 100 mq (0.3 mmol) with stirring. Heat the solution to 100° for 2.5 hours. T:c,c
The reaction is followed by acetone/cyclohexane (son acetone/cyclohexane) and heating is terminated when the conversion is approximately 75% complete (by phosphorus/phosphoric acid spray). The cooled solution is
N a+: Pour into t(1oo*), 20,j KtO
Extract with Ac. Wash the separated organic phase with an additional 50 g of water. The aqueous phases are combined and back extracted with 20 d of EtOAc. The organic phases were then combined, dried over Na2So4, concentrated in vacuo and placed on a column of silica gel 1007.
Dissolve with 50% acetone/cyclohexane. This yields 45Wli of 4,5-pyrrolo-6-hydroxyindoline (U-62,370) with 25η of starting material and +10 of raw material (isolated yield of 44%, based on recovered starting material of 69%).

NMR (ACet-d,) + 7.8(br, B
, IH), 7.03 (s, IH)6.83 (
8. 1 H), 4.25 - 3.25 (
m, 5E[). 2.86 (8, 3H), 2.
36 (S,3H) The product is treated with 1.0-acetic anhydride and 20 η of NaOAc overnight and taken up in CH2Ct2/H20.

The organic phase is separated, dried on ua2so, and concentrated.

NMR (cpcz3): 7,8(br-fi
l, ta), 7. +6 (s+IH), 6.97 (
il, L H ), 4.42, 4.20 (
(La, 2H), 4.2-3.7 (m, 3H),
2.86(e, 3H). 2.40 (8.3H)
, 2.35 (al3H), 2.06 (θ.3H) u: m/e M+380 (25%). 199
(100%)-2'-1% SE-+ This compound was assayed against L 1210 Mus musculus leukemia cells in culture by a standard test tube dilution assay;
I got the following results.

D, o (μg/+ug) =)5 D90 (μ
g/+d) => 5 亙dan'' 4.5-Hiroro 6-
4. From human 9 roxyindoline alcohol exclamation. Preparation of s-Hirolow 6-hydroxyindoline bromide (11) Alcohol base in 1.0 ml of dry acetonitrile 25 η
(65 micromoles) in CBr, 33η under nitrogen at room temperature.
(100 μmol) and human rifenylphosphine (P
h, P ) 26η (100 micromol) is added. After cutting the blade 1, add additional CBr, 11 # and Ph3
Add P8rNi. After a total of ω minutes, the reaction was changed to CH2Ct2/
It will be taken up during H20. Separate the organic phase and add Na, So,
Dry on top and concentrate. 20 x 20 (250 m
Place the residue on 3 μ-silica dull plates and elute with 50% acetone/cyclohexane. Products with high Rf values (
0.64; Rf of alcohol==0.45), i.e. 4,5-pyrrolo-6 hydroxyindoline bromide (U-62.694') of the compound till about 8
Collect ■.

NMR (CDCt3): 8.5(1)r. 8,
LH), 7.1 (S, IH), 6.9 ('LIH
L4.23 (d+2H+J=6Hz) l4-0 3
-5 (m+3H) ,2.89(s,3H),
2.38 (S, 3H).

Peilstein test: Positive MS: C16Hz3N203” for BrSSi,
Calculated value 430.0382 Measured value 430.0375 (
m/-TMS); m/eM+430/432 (
14%), 271 (90%),! 47 (100%
).

This compound was tested in a standard test tube dilution assay and
The assay was performed on 1210 Mus musculus leukemia cells and the following results were obtained.

I'sa (μg/m/) = 0.12 Dso (
μg/+d) = o. 37 Example 4 4.5 - Preparation of 415 - Hirolo = 6-hydroxyindoline mesylate uυ from Hirolo 6-hydroxyindoline alcohol α0 Alcohol substrate (10) in pyridine 1.0 20
mg (63 micrommol) in a water bath! Add 8 μt (70 micromoles) of methanesulfonyl chloride under N2 while stirring. After the addition, an additional 2ttL of CH, So2Ct was added, and after a total reaction time of ω minutes, 2N HCz/
Finish with CH2Ct2.

C) It shows a somewhat high Rf value (0.66').

Separation TLC (20x2) (m, 3 250μ silica plates) uses a high Rf material (NMR pattern with only one CH).
3802 units.

Indicated. 2 mq and low Rf material i.e. the compound aυ (U-62,695
) produces 9 mq.

NMR (Acet.-a,): 8.6 (br.
s, I H), 6, g7 (s, I H), 6
．． 74 (511 1H). 4.3 (m, 2H)
．． 4.1-3.6 (m. 3 H). 2.96
(8.3H), 2.79 (8.3EL),
2.26 (B. 3H).

The compound was assayed against L 1210 Mus musculus leukemia cells in culture by a standard in vitro dilution assay;
I got the following results.

D=o (μg,/m0= 1.0 D20(
μq/m0=3.3f1k1 Example 7 i4+8+sa-
Cycloderopa [c] Henso (1.2-b: 4.3
-b') diviror-4(5H) one (13,N-
Preparation of (methylsulfonyl)-4. Follow the procedure to make 5-hyloe+-6-hydroxyindoline bromide (step 10) but with a high Rf (0
．． If the reaction mixture before finishing the product bromide of 64') is compressed in vacuo and applied directly to the silica glue plate of the polishing layer, a new low R
A band of f (0.32) was observed and recovered as compound α3.

NMR (CDCts): 9-5 (br.8
,1 ”) ,6 .83'("r"a),6
．． 34 (s, Hb), 4.10 ((L.
Hc ), 3.93 (dd, Hd), 3.04 (sl
3H) l2.93 (m,”e)+2.00 (
d, 3H), 1.97 (coal Hf)
, 1.37 (act, I{g).

JC8=0.0 eclipse Je, g=4.4JC, (L=
9.7 Jf,g=4.4Jd,e ” 4
．． 7 J NH,a == 2, OJ
e,f=7.7 Ja,CH,=<1.0M
S: Lusolylated pam/e M+ 386 with BSTFA (DMF containing 1% TMS-Ct)
/388 (22, corresponding to product + Me3SiCL) yielded 12%.

Uv: (methanol) λ 224, 272, 338 This compound was tested in a standard test tube dilution assay at L 1 of the culture medium.
210 Mus musculus leukemia cells and obtained the following results.

Engineering D5o (μg/rnt)=0. l3 Engineering D9o (μ
g/wg = 0.4297 kg Compound (1 Alternative Preparation 4.5-pyrrolo6 in methylene chloride lrnt
-Hydroxyindolinebromide (or mesylate)
(0.1 mmol) of diisodepyruedylamine O
. Add 1 mmol and stir at room temperature under N2 for 24 hours. The reaction solution was taken up in 10 d of methylene chloride, and 0,1
Wash with NHCt, Ha2So. Drying and concentration above yields the desired product. silica chromatography 7
It can be further refined using

Compounds aυ and (1) exhibit anti-madder activity against Bacillus subtilis, Klebsiella pneumoniae, S. lutea, Staphylococcus aureus, and M. avium.

Formula ゝ l ↓Stage 9 αυ ↓Stage 1l N 2 (1) (2) ↓Stage 2 (4) (3) ↓Stage 4 (5} (6) ↓Stage 6 (8) (7) Only definition, == CH.3-, -CH2Ph, CH2'=C
HCH2-, -CH2SCH3, -CH20CH3, -
cH2ocH2CH20CH3, -CH2CCI,?
s), phenyl [Note = R3 is 0- with respect to compound (2)
Alkyl only] R, = So2R2, So CH2COPh, Co
2CH2Z! == So2R2, C1, Br, IY
= Li, Na, K, MgXZ = CH2 Engineering, CCI3, CH2SO2R2, Ph, 7 Luo L
/ Nylmethyl As used herein, alkyl of 1 to 2 or 1 to 5 carbon atoms includes methyl, ethyl, proyl, butyl, intyl, and branched chain isomers thereof.

Expression 3 Expression 4 Reference example 1 (2) +3+ U-62,598 {3) (4) U-62,597 R example 3 (4) (5) U-62,586 Temple forgery example 4 (5) (6) Guisei line 5 Example 3 (6) (7) U-62,696 01 (Lυ U-62,694 Example cow {7} (8) U 62,697 Example 1 αO αD U-62,695 Reference example 7 (8) (9) U-62,233 Example 2 αυ 0 U-62,736 (9) 00 σ-92,370

Claims (1)

[Claims] 1. Compound of the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ [In the formula, R_1 is H, CH_3-, -CH_2Ph, CH
_2=CHCH_2-, -CH_2SCH_3, -CH
_2OCH_3, -CH_2OCH_2CH_2OCH
_3, -CH_2CCl_3, and -CH_2CH_2
selected from the group consisting of Si(R_2)_3, R_2, R
_2′ and R_3 are hydrogen, alkyl of 1 to 5 carbon atoms, or phenyl, and R_4 is SO_2R_2, S
selected from the group consisting of O_2CH_2CO phenyl, CO_2CH_2Z, and X is OSO_2R_2, Cl, Br
, l and OH]. 2. A compound of the formula ▲ There are mathematical formulas, chemical formulas, tables, etc. The formula consists of reacting with a reagent selected from the group consisting of triphenylphosphine, carbon tetrabromide/triphenylphosphine and N-iodosuccinimide/triphenylphosphine. R_2, R_2', R_3 and R_4 are as defined above, and X is -OSO_2R_2, Cl, B
selected from the group consisting of r and l. 3. There are formulas, chemical formulas, tables, etc., according to claim 2, which consists of reacting the compound of formula ▲ with triphenylphosphine/carbon tetrabromide. ▼ Method for producing the compound.