JPH0465076B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The antibiotic CC-1065 is characterized by chemical and physical variables and is claimed in US Pat. No. 4,169,888. After that, the structure of antibiotic CC-1065 was published as "Structure of antibiotic CC-1065".
"Proof of Amtibiotic CC-1065)" DG Martin, CG Chidester, DJ Duchamp, and SAMizsak, J. Antibiot.33 It was clarified as revealed in Volume 902 (1980).
The structure of antibiotic CC-1065 is shown in Formula 1. Antibiotic CC-1065 consists of a system of three fragments, the most unstable part of the molecule being 1,2,8,8a-cyclopropa[C]benzo[1,2-b:4,3-b']dipyrrole. -4(5H)-one, herein designated as compound (12). Antibiotic CC-
Attempts to obtain this fragment by degradation of 1065 were unsuccessful. For this reason, no prior art methods for obtaining compound (12) are known.

produced from the compounds (9), (10), (11) of the present invention,
Compound (12) is made by the 11-step method revealed in Formula 2. Compound (12) and this compound (9)
~(11), as well as certain intermediates disclosed herein, are compatible with certain bacteria, such as Bacillus subtilis.
subtilis), Klebsiella pneumoniae, Sarcina lutea, Staphylococcus aureus, and Mycobacterium avium.
avium) active against bacteria. These compounds can therefore be used to disinfect washed and stacked tableware contaminated with Staphylococcus aureus. Furthermore, the antimicrobially active compounds of the present invention can be used as a bacteriostatic rinse agent for laundered clothing, as a bacteriostatic rinse agent for impregnated paper and fabrics, and to inhibit the growth of susceptible bacteria in plate assays and microbial culture media. is also useful. In general, the antimicrobially active compounds of the present invention can be used in the same manner as disclosed for antibiotic CC-1065 in US Pat. No. 4,169,888. 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, the 11 step method for making compound (12) is shown in Formula 2. This stage is as follows.

Step 1 - The first step of the synthetic clue (nucleophilic substitution of aromatics) was developed by G. Bourdais (J.
Bourdais) and C. Mayer (C.
Mahieu), Compt. Redux [C], vol. 263, 84
(1966). Also, G. Bourdais and C. Germain, Tet. Letters 195
(1970). The various R ₁ groups can be synthesized (1) by procedures described in the literature (appropriate references detailed under Step 8).
phenolic precursors.
The various malonates, β-ketoesters and β-diketones used are all known compounds.

Step 2 - Reduction. When R′ ₂ =alkoxy, diisobutylaluminum hydride is the reagent of choice. The reaction conditions are very specific for optimal yields (Reference Example 1)
). When R′ ₂ =alkyl or phenyl, standard reduction procedures using sodium borohydride can be used.

Step 3 - Exchange of functional groups. _In particular, the chemistry described herein is for 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. The halogen analogues of No. 4 are Ph ₃ P/CCl ₄ (CBr ₄ ) and N
- can be made by standard procedures known in the art such as iodosuccinimide/triphenylphosphine.

Step 4 - Reduction - Cyclization. This step is a new method for preparing indolines (dihydroindoles). This involves reduction of the nitro to amino group and concomitant intramolecular cyclization, yielding (5). The reduction step detailed herein utilizes H ₂ , Pto ₂ in an alcohol in the presence of a tertiary amine. These are 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. Alternative reducing situations can use Fe or Ticl ₃ or Sncl ₂ in acid. This would then require another step involving base treatment to cause cyclization to (5). An example of reduction with iron is Fe/CH ₃ CO ₂
Use H/CH ₃ CH ₂ OH (G.S.
GS Ponticello and JJ Baldwin
Baldwin), J.Org.Chem. vol. 44, p. 4003 (1979)). R ₁ = CH ₂ Ph or −CH ₂ CH
In the case of =CH ₂ , these conditions are necessary.

The idea of nitro reduction followed by in situ cyclization to indole was proposed by A.D. Batcho.
and W. Leim-gruber, DE 2057840 (1971), and is a significant improvement over the old Reissert procedure of reductive cyclization to indoles. Law (R.G.A.
RJ Sundberg, The Chemistry of Indoles
Indoles), pp. 176-183, Academic
Press, New York, 1970).

Step 5 - Substitution of unstable groups. This stage is stage 8
This is necessary because X is incompatible in the chemical reaction of This consists of replacing X with an acetate or a conjugate base of a _C1 - _C5 alkyl carboxylic acid under standard conditions (alkali carboxylate in acetone, DMF or alcohol).
Since some hydrolysis may also occur when X=SO ₂ CH ₃ , the reaction mixture is treated with acetic anhydride before isolation of (6).

Step 6 - Nitration can be carried out under a variety of conditions described in the literature including nitric acid in acetic acid, acetic anhydride, sulfuric acid, acetic acid/ _H2O , alcohols and nitroalkanes. The region selectivity of this reaction is supported by the spectroscopic data obtained.

Step 7 - Reduction of nitro to amino group is performed in Step 4
Follow the same chemical reaction description, but omit the base.

Step 8 - Indole synthesis. This procedure is outlined in Gassman's Indole Chemistry [PG Cassman et al., J.Am.Chem.
Soc. 96 pages 5494, 5508, 5512 (1974
year)]. 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. The present method deviates from the published Gassmann route by using chlorosulfonium complex A and reacting it with aniline 6. Gassmann created chloroamines of aniline and reacted them with thioethers to create oxindoles. Second, the method uses two different chlorines, whereas Gasman uses 2 equivalents of aniline followed by 2 chlorine. Triethylamine, diisopropylethylamine, bis(1,8-dimethylamino)naphthalene, etc. work well as both base 1 and base 2, but bis(1,8-dimethylamine)naphthalene is preferred as base 1; Base 2 is triethylamine. A variety of solvents can be used, such as chloroform, acetonitrile, tetrahydrofuran (THF) and methylene chloride, with the latter being preferred. Temperature range is -50 to -80°
The reaction is carried out under an inert atmosphere. Cyclization to oxindole B is best promoted by acid catalysis (2N HCl, ether and/or ethyl acetate) as described by Gassmann.

The final reduction (reductive removal) to (9) can be achieved with lithium aluminum hydride (as described by Gassmann) or diborane type reagents, the latter being by far superior. The preferred reagent is 24 hours at room temperature.
It is (CH ₃ ) ₂ S·BH ₃ in THF.

Step 9 - This deprotection step (removal of _R1 ) is described in detail in Example 8 for _R1 = _CH3 . Many procedures have been described in this technique, including the cleavage of methyl ether;
Hexamethylphosphoric triamide (HMPA) under inert atmosphere (95-100°)
Only the alkyl mercaptide among them was found to be effective [SC Welch (SC Welch)]
Welch) and A.S.C.P.
Rao (ASCPRao), Tet. Letters 505
No. (1977) and TRKelly, HMDali and WGTsang, Tet. Letters, 3859
(1977), or in dichloroethane.
Me ₂ S. BBr ₃ (PG Willard and CBFryhle, Tet. Letters No. 3731 (1980)).

When R ₁ = CH ₂ Ph, standard hydrogenolysis conditions (H ₂ , Pd/C) are fully suitable for deprotection [Org.Reactions Vol. 7
263 pages (1953)]. When R ₁ = CH ₂ SCH ₃ mercuric chloride in acetonitrile/H ₂ O removes the ether (RA Holton and RG Davis).
Davis), Tet. Letters No. 533 (1977)].
When R ₁ = CH ₂ OCH ₃ , a mild acid such as acetic acid produces phenol 10 [J.
Med.Chem. Volume 9, page 1 (1966) or
Synthesis 244 (1976)]. However, in reality, this protecting group is lost in step 6;
Can be reintroduced prior to stage 7 under standard conditions. When R ₁ =CH ₂ OCH ₂ CH ₂ OCH ₃ , phenol can be made from ZnBr ₂ or TiCl ₄ in CH ₂ Cl ₂ [Tet. Letters No. 809 (1976)]. When R ₁ =-CH ₂ CH=CH ₂ , several two-step procedures carry out the deprotection of the ether [Pd in alcohol/
C, Ang.Chem.Int.Ed. vol. 15, p. 558 (1976)
year); SeO ₂ , CH ₂ CO ₃ in dioxane
H, Tet. Letters No. 2885 (1970);
−BuOK, DMSO followed by H ₂ SO ₄ in acetone, J.Chem.Soc.1903 (1965);
RhCl( _PPh3 ) ₃ in alcohol,
DABCO followed by pH2, J.Org.Chem.38
Volume 3224 (1973)]. R ₁ =−CH ₂ CH ₂ Si
When (R ₂ ) ₃ , deprotection is carried out with Bu ₄ NF [H.
Gerlach) et al., Helv.Chim.Acta.60 volumes
3039 pages (1977)].

Step 10 - See Step 3.

Step 11 - This step (when X=Br) is facilitated by contact with silica gel and also occurs on standing in a protic solvent. This reaction also proceeds in the presence of aqueous bases such as tertiary amines, pyridine, t-butoxide, and weak bases such as bicarbonates and carbonates.

The following examples are illustrative of the production arrangements and methods of the present 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.

Reference example 1 Aryl diethyl malonate 2 to 2
Preparation of -aryl-1,3-propanediol 3 100 g (0.07 mol) of diisobutylaluminum hydride (DIBAL) in 400 ml of toluene are added to 400 ml of THF in _N2 cooled in an ice-water bath. To this stirred solution is added 33.0 g (0.105 mol) of aryl malonate 2 in 100 ml of THF. The addition rate is adjusted to keep the reaction temperature above 5°. After the addition is complete, remove the ice bath. After a total reaction time of 3 hours, the reaction is stopped by adding the solution in small portions to cold 3N HCl (ca. 1.5) with stirring. The mixture is then extracted with 1 portion of EtOAc followed by 1000 ml of CH ₂ Cl ₂ . The combined organic phases are dried over Na ₂ SO ₄ and concentrated to a reddish-brown residue (21.2 g). The residue was chromatographed on 500 g of silica gel with a gradient of 60% EtOAc/hexanes → 100% EtOAc to give 11.7 g of diol 3 (U-62598).
(49% yield) as a pale red oil (will solidify if left in the freezer).

NMR ( _CDCl3 ): 7.5-7.0 (m, 3H), 3.80 (s,
3H), 4.0−3.3 (m, including 7H−2OH).

MS.C ₁₀ H ₁₃ NO ₅ Calculated value: 227.0794 Measured value: 227.0780 Analysis: Calculated value: C, 52.86, H, 5.76, N, 6.16 Measured value: C, 53.40, H, 5.77, N, 5.99 Reference example 2 2 - Preparation of 2-aryl-1,3-propanediol bismesylate 4 from aryl-1,3-propanediol 3 4.7 g (0.2 mol) of diol 3 in 100 ml of dry pyridine at 0-5° under _N2 Add 6.8 g (0.06 mol) of methanesulfonyl chloride while stirring. at 5°
After stirring for 30 minutes and 90 minutes at room temperature, the solution is concentrated in vacuo and taken up in CH ₂ Cl ₂ /1N HCl. The organic phase is separated, dried over Na ₂ SO ₄ and concentrated to a residue. Trituration with EtOAc yields an off-white solid, and the mother liquor is converted into silica gel (EtOAc).
Compound (4) bismesylate (U-
62597). NMR (Acet-d ₆ ): 7.7-7.2 (m, 3H), 4.62 (d,
4H, J=JHz), 4.11(t, 1H,
J=7Hz), 3.92 (s, 3H), 3.06
(s, 6H).

Analysis: For C ₁₂ H ₁₇ NO ₉ S ₂ , Calculated value C, 37.59, H, 4.47, N, 3.65 Measured value C, 37.35, H, 4.44, N, 3.59 This compound was applied to L1210 Mus musculus leukemia cells in culture medium. The following results were obtained using a standard test tube dilution assay.

LD ₅₀ (μg/ml) = 6.0 ID ₉₀ (μg/ml) = 18 Reference Example 3 Preparation of 6-methoxyindoline bismesylate 5 from 2-aryl-1,3-propanediol bismesylate 4 THF 30ml: EtOAc 20ml , and absolute ethanol
1.5 ml of triethylamine and 400 mg of PtO ₂ are added to 1.91 g (0.005 mol) of compound (4) in 150 ml. While shaking this solution, the hydrogen pressure was 0.492-0.703Kg/cm ² (7
-10psi) for 30 minutes. The reaction solution is then filtered over Celite and concentrated in vacuo. several times in vacuum
Azeotrope with CH ₂ Cl ₂ and then convert the residue into CH ₂ Cl ₂ 100
ml and cool under _N2 in an ice bath. Add 1.5 ml of triethylamine to the stirred solution.
is added followed by the dropwise addition of 900μ of methanesulfonyl chloride. After stirring for 30 minutes, allow the solution to reach room temperature for 60 minutes. Then wash with solution 1N HCl,
Dry _{over Na2SO4} and concentrate. The residue was rapidly chromatographed on 150 g of silica gel with 500 ml of 60% EtOAc/hexane followed by 1000 ml of 80% eluent as an off-white solid, mp 122~
Compound (5), 6- at 123° (recrystallized from ethanol)
Methoxyindoline bismesylate (U-62586)
1.3g (78% yield) was recovered. NMR (DMF- _d7 ): 7.36 (d, 1H, J=8.5Hz),
7.00 (d, 1H, J=2Hz), 6.69
(dd, 1H, J=2, 8.5Hz), 4.47
(2H, d, J=6Hz), 4.3−3.6
(m, 3H), 3.80 (s, 3H), 3.20
(s, 3H), 3.07 (s, 3H).

Analysis: For C ₁₂ H ₁₇ NS ₂ O ₆ Calculated C, 42.97, H, 5.11, N, 4.18 Measured C, 42.87, H, 5.27, N, 4.29 This compound was tested using a standard test tube dilution assay. ,
The culture medium was assayed against L1210 Mus musculus leukemia cells, and the following results were obtained.

ID ₅₀ (μg/ml) = 4.8 ID ₉₀ (μg/ml) = 10 Reference Example 4 Preparation of 6-methoxyindoline bismesylate 6 from 6-methoxyindoline bismesylate 5 6-methoxyindoline bismesylate 5 in 30 ml DMF 13.0 g (39 mmol) are added with 800 ml of absolute ethanol followed by 32 g of sodium acetate. This heterogeneous mixture was refluxed under _N2 for 24 h and
Cool and concentrate in vacuo. Dilute the residue with acetic anhydride
Treat with 100ml for 2 hours (while stirring at room temperature),
Then concentrate in vacuo. The residue was dissolved in CH ₂ Cl ₂ /H ₂ O.
Separate the organic phase, dry over Na ₂ SO ₄ , filter through charcoal and concentrate to an oil. The oil solidifies to yield 11.6 g of compound (6), 6-methoxyindoline acetate (100%, 60% if necessary).
Further purification is possible by silica gel chromatography using EtOAc/hexane eluent). NMR ( _CDCl3 ): 7.17 (d, 1H, J=8.5Hz),
7.02 (d, 1H, J=2Hz), 6.60
(dd, 1H, J=2, 8.5Hz), 4.18
(d, 2H, J=6Hz), 4.1−3.4
(m, 3H), 3.78 (s, 3H), 2.91
(s, 3H), 2.05 (s, 3H).

Analysis For C ₁₃ H ₁₇ NO ₅ S, Calculated value: C, 52.16, H, 5.76, N, 4.68 Measured value: C, 52.13, H, 5.79, N, 5.27 MS : Calculated value: 299.0827 Measured value: 299.0823 Reference Example 5 6-methoxyindoline acetate 6
Preparation of 5-nitro-6-methoxyindoline acetate 7 from 500 ml (1.67 mmol) of 6-methoxyindoline acetate 6 in 20 ml of nitromethane at 90%
Add 90μ of _HNO3 . Stir the cooled (0-5°) reaction solution for 30 minutes, then warm to room temperature for 30 minutes.
The solution is diluted with CH ₂ Cl ₂ and aqueous sodium bicarbonate. Separate _the organic phase, dry over _Na2SO4 ,
Concentrate. The residue was chromatographed on 50 g of silica gel (60% EtOAc/hexane eluent → 100%
EtOAc) gives a yellow solid, melting point 175-177°
(recrystallized from ethanol), compound (7), 5-nitro-6-methoxyindoline acetate (U-
62696) yields 440 mg (76% yield). NMR (DMF- _d7 ): 7.91 (s, 1H), 7.20 (s,
1H), 4.27 (d, 2H, J=6Hz),
4.3−3.7 (m, 3H), 3.98 (s,
3H), 3.17 (s, 3H), 2.07 (s,
3H).

Analysis: For C ₁₃ H ₁₆ N ₂ O ₇ S, Calculated value: C, 45.34, H, 4.68, N, 8.14 Measured value: C, 44.81, H, 4.77, N, 8.16 This compound was placed in a standard test tube. By dilution assay,
It was assayed against L1210 Mus musculus leukemia cells in culture and the following results were obtained.

ID ₅₀ (μg/ml) =>50 ID ₉₀ (μg/ml) =>
50 Reference Example 6 Preparation of 5-amino-6-methoxyindoline acetate 8 from 5-nitro-6-methoxyindoline acetate 7 5-Nitro-6-methoxyindoline acetate 7 in 50 ml THF and 150 ml absolute ethanol
Add 500 mg of PtO ₂ to 4.5 g (13 mmol) and add H ₂
below 0.703Kg/cm ² (10psi) until ingestion stops (approx.
60 minutes) Shake. Filter and concentrate in vacuo.
After concentration, 3.0 g of product precipitates out. Filter this out,
Rapid chromatography of the mother liquor on 100 g of silica gel with EtOAc eluent yields an additional 0.6 g. Total yield (3.6) of 5-amino-6-methoxyindoline acetate (U-62697) of compound (8)
g) is 88%, melting point 134-135°, (from acetone/cyclohexane). NMR (CDCl ₃ ): 7.02 (s, 1H), 6.65 (s, 1H),
4.16 (d, 2H, J=6Hz), 4.1−
3.5 (m, 3H), 3.83 (s, 3H),
3.6 (br.s, 2H), 2.83 (s, 3H).

Analysis: For C ₁₃ H ₁₈ N ₂ O ₅ S Calculated: C, 49.67, H, 5.77, N, 8.91 Measured: C, 49.74, H, 5.72, N, 8.94 This compound was diluted in a standard test tube. By the test,
It was assayed against L1210 Mus musculus leukemia cells in culture and the following results were obtained.

ID ₅₀ (μg/ml) =>50 ID ₉₀ (μg/ml) =>
50 Example 1 5-amino-6-methoxyindoline acetate 8 to 4,5-pyrrolo-6-
Preparation of methoxyindoline 9 _Cl2 / _CH3 in 14 ml _{of CH2Cl2} dried at -75° under nitrogen
Add 6.0 ml of Cl ₂ solution (20 μ Cl ₂ /ml CH ₂ Cl ₂ ). Add CH ₃ (CH ₃ S) CHCO ₂ to this stirred solution.
C ₂ H ₅ 370 μ (2.5 mmol) [EH Wick, T.
Yamanishi), H.
C.Wertheimer), Y.E.
Hoff), BF Proctor
and SA Gold Bliss (SA
Goldblith), J.Agr.Food Chem. vol. 9, p. 289 (1961
CH ₃ (Br)CHCO ₂ C ₂ H ₅ and methyl mercaptide] is added according to the procedure of 2010). 5 minutes later,
A solution of 470 ml ( _2.2 mmol) of 1,8-bisdimethylaminonaphthalene and 628 mg (2.0 mmol) of anilinoindoline 8 in 3.0 ml of dry CH ₂ Cl 2 is added dropwise over 15 minutes. Stir the red solution at −75° for 2 hours to prepare triethylamine in 650μ of CH ₂ Cl ₂
Add 350μ dropwise over several minutes. Remove cooling bath.
When the reaction solution reaches room temperature, it is briefly concentrated in vacuo. Add 5 ml of EtOAc to the residue and 25 ml of ether.
ml and 6 ml of 2N HCl and stir vigorously for 2 hours. The organic phase was separated and the aqueous layer was dissolved in EtOAc/ _Et2O .
(1:1) extraction. Combine the organic layers and add _Na2
Dry over _SO4 and concentrate. At this point the residue
Take up in 10 ml of THF and treat with 3.0 ml of 2M BH ₃ .SMe ₂ overnight at room temperature under nitrogen. Alternatively, the diastereomeric oxindole B derived from acid treatment can be isolated at this point by silica gel chromatography (50 g, 60% EtOAc/hexanes to 90% EtOAc/hexanes). GC-MS : m/e M ⁺ 414 (15%), 227 (100%)
−2′1%SE−30 NMR (CDCl ₃ ): 8.4 (br.s, 1H), 7.11 (s, 1H),
4.5−3.7 (m, 5H), 3.90 (s,
3H), 2.95 (s, 3H), 2.10 (s,
3H), 1.92 (s, 3H), 1.82 (s,
3H) - major diastereomer (2.5/1); minor diastereomer shows the following differences: −SCH ₃
and −CH ₃ is 1.99(s,
3H), 1.76 (s, 3H), NH is
7.7, _CH2 band is at 4.3-3.6ppm.

The aqueous phase from acid treatment is neutralized and extracted _{with CH2Cl2} . _{The CH2Cl2} solution was dried, concentrated and chromatographed on silica gel with 50% acetone/cyclohexane resulting in 40% recovery of starting material (anilinoindoline) and deacylated starting material.
yields 20%.

The borane dimethyl sulfide reductive removal reaction (to B) is worked up by quenching with 1N HCl until gas evolution ceases and taken up in CH ₂ Cl ₂ /H ₂ O. The separated organic phase is dried over Na ₂ SO ₄ and concentrated. Chromatography of the residue on silica gel (50% acetone/cyclohexane) yields 155 mg of product (9). Isolated yield 25%, 85% based on recovered starting material. Melting point 182~
183° (phase change at 160°, recrystallized from chloroform).
Product 4,5-pyrrolo-6-methoxyindoline
(9) (U-62, 233). NMR (CDCl ₃ ): 8.3 (br.s, 1H), 6.96 (s, 2H),
4.2−3.5 (m, 5H+0H), 3.92
(s, 3H), 2.87 (s, 3H), 2.41
(s, 3H).

Analysis For C ₁₄ H ₁₈ N ₂ O ₄ S Calculated value: C, 54.17, H, 5.84, N, 9.03 Measured value: C, 53.49, H, 5.96, N, 9.42 GC-MS: O-acetate-m /eM ⁺ 352(13
%), 213 (100%) −2′−1%SE−
30, temperature 150-260° (10°/min), single peak.

This compound was tested using a standard test tube dilution assay.
The culture medium was assayed against L1210 Mus musculus leukemia cells, and the following results were obtained.

ID ₅₀ (μg/ml) =>4 ID ₉₀ (μg/ml) =>
4 Example 2 Preparation of 4,5-pyrrolo-6-hydroxyindoline 10 from 4,5-pyrrolo-6-methoxyindoline 9 To 10 ml of dried and degassed HMPA (hexamethylphosphoric acid triamide) was added 350μ of butyl mercaptan at room temperature. Add. Cool the solution in an ice water bath;
2.0 ml of 1.5Mn-BuLi in hexane are added dropwise. After cooling the reaction to room temperature, add 100 mg of indole 9.
(0.3 mmol) while stirring. solution
Heat to 100° for 2.5 hours. The reaction was followed by TLC (50% acetone/cyclohexane) and the conversion was approx.
When it reaches 75% completion (by vanillin/phosphoric acid spray), stop heating. Add cooled solution to 1N HCl
(100 ml) and extract with 20 ml EtOAc. Wash the separated organic phase with an additional 50 ml of water. The aqueous phases are combined and back extracted with 20 ml of EtOAc. The organic phases are then combined, dried over Na ₂ SO ₄ , concentrated in vacuo, loaded onto a 100 g column of silica gel and eluted with 50% acetone/cyclohexane. Starting material 25mg
yielding 45 mg of 4,5-pyrrolo-6-hydroxyindoline (U-62370) of product 10 (isolated yield
44%, 69% based on recovered starting material). NMR (Acet-d ₆ ): 7.8 (br.s, 1H), 7.03 (s,
1H), 6.83 (s, 1H), 4.25−3.25
(m, 5H), 2.86 (s, 3H), 2.36
(s, 3H).

The product is treated with 1.0 ml of acetic anhydride and 20 mg of NaOAc overnight and taken up in CH ₂ Cl ₂ /H ₂ O. Separate the organic phase, dry over Na ₂ SO ₄ and concentrate. NMR (CDCl ₃ ): 7.8 (br.s, 1H), 7.16 (s, 1H),
6.97 (s, 1H), 4.42, 4.20 (dd,
2H), 4.2-3.7 (m, 3H), 2.86
(s, 3H), 2.40 (s, 3H), 2.35
(s, 3H), 2.06 (s, 3H). GC-MS : m/eM ⁺ 380 (25%), 199 (100%) −
2'-1%SE-30 This compound was tested using a standard test tube dilution assay.
The culture medium was assayed against L1210 Mus musculus leukemia cells, and the following results were obtained.

ID ₅₀ (μg/ml) =>5 ID ₉₀ (μg/ml) =>
5 Example 3 4,5-pyrrolo-6-hydroxyindoline alcohol 10 to 4,5-pyrrolo-6-hydroxyindoline bromide
Preparation of 11 alcohol substrate in 1.0 ml of dry acetonitrile
25 mg (65 micromol) of _CBr4 under nitrogen at room temperature
Add 33 mg (100 μmol) and 26 mg (100 μmol) of triphenylphosphine (Ph ₃ P). Stir for 30 minutes, then add an additional 11 mg of CBr ₄ and 8 mg of Ph ₃ P. After a total of 60 minutes, the reaction is taken up in CH ₂ Cl ₂ /H ₂ O. Separate the organic phase, dry over Na ₂ SO ₄ and concentrate. 20×20cm 250μ silica gel plate 3
Place the residue on a plate and elute with 50% acetone/cyclohexane. About 8 mg of a product with a high Rf value (0.64; Rf of alcohol = 0.45), 4,5-pyrrolo-6-hydroxyindoline bromide (U-62694) of compound (11), is recovered.

NMR (CDCl ₃ ): 8.5 (br.s, 1H), 7.1 (s, 1H),
6.9 (s, 1H), 4.23 (d, 2H, J
=6Hz), 4.0−3.5(m, 3H),
2.89 (s, 3H), 2.38 (s, 3H).

Weilstein Test: Positive MS: Calculated value 430.0382 Measured value 430.0375 (Mono TMS) for C ₁₆ H ₂₃ N ₂ O ₃ 79BrSSi; m/eM ⁺ 430/432 (14%), 271
(90%), 147 (100%).

This compound was assayed against L1210 Mus musculus leukemia cells in culture by a standard in vitro dilution assay with the following results.

ID ₅₀ (μg/ml) = 0.12 ID ₉₀ (μg/ml) =
0.37 Example 4 4,5-pyrrolo-6-hydroxyindoline alcohol 10 to 4,5-pyrrolo-6-hydroxyindoline mesylate
Preparation of 11 20 mg of alcohol substrate 10 in 1.0 ml of pyridine (65
micromillimol) while stirring in an ice bath.
Add 8 µm (70 µmol) of methanesulfonyl chloride under _N2 . After 30 minutes, add an additional 2μ of CH ₃ SO ₂ Cl and work up with 2N HCl/CH ₂ Cl ₂ after a total reaction time of 60 minutes. Separate the organic phase, dry over Na ₂ SO ₄ and concentrate. TLC shows mostly low Rf product (0.28 in 50% acetone/cyclohexane, alcohol Rf = 0.46) and some high Rf product (0.66). Separation TLC (20×20
cm, three 250μ silica gel plates), 2 mg of high Rf material (NMR showed only one CH ₃ SO ₂ group, probably this is chloride) and 2 mg of low Rf material i.e. compound (11) (U- 62695) yields 9 mg. NMR (Acet-d ₆ ): 8.6 (br.s, 1H), 6.97 (s,
1H), 6.74 (s, 1H), 4.3 (m,
2H), 4.1-3.6 (m, 3H), 2.96
(s, 3H), 2.79 (s, 3H), 2.26
(s, 3H).

This compound was tested by standard test tube dilution assay.
The culture medium was assayed against L1210 Mus musculus leukemia cells, and the following results were obtained.

ID ₅₀ (μg/ml) = 1.0 ID ₉₀ (μg/ml) = 3.3 Example 5 Step a 6-(benzyloxy)-2,3-dihydro-1-(methylsulfonyl)-5-nitro-1H-indole -3-Methanol acetate 7 6-(benzyloxy)- in nitric acid (90%, 4.4 ml, 94 mmol) and dry nitromethane 11
2,3-dihydro-1-(methylsulfonyl)-5
Added to a solution of -nitro-1H-indole-3-methanol acetate (24.6 g, 65.4 mmol) at 0°C under nitrogen. After stirring for 40 minutes, the reaction mixture was diluted with methylene chloride, 5% NaOH and saturated NaCl.
and dried (Na ₂ SO ₄ ). Evaporative crystallization from ethanol left a brown solid; further recrystallization from acetic acid and ethanol yielded 17.7 g (64%
The title compound (yield) was obtained. Melting point 145−147
℃. NMR (CDCl) δ 2.07 (s, 3H), 2.82 (s,
3H), 3.40-4.40 (m, 5H), 5.29 (s, 2H), 7.23
(s, 1H), 7.3-7.67 (m, 5H), 7.88 (s, 1H).

Step b 5-amino-6-(benzyloxy)-2,
3-dihydro-1-(methylsulfonyl)
-1H-indole-3-methanol acetate 8 Hydrogenation of the compound of step a (20 g, 48 mmol) in THF (190 ml) and ethanol (530 ml)
40psi on _PtO2 (84.5% catalyst, 1.8g, 6.7mmol)
It lasted two hours. The catalyst was filtered off and washed with ethanol. The product crystallized from the cooled filtrate. Chromatography of the mother liquor on silica gel eluting with ethyl acetate and hexane gave additional title compound. (Total yield 15g, 81
%) Melting point 132-134: ClR (undiluted) 3425, 5350,
2910, 1720, 1605, 1490, 1330, 1230, ^{1060cm -
1} ; NMR (CDCl ₃ ), 82.06 (s, 3H), 2.74 (s,
3H), 3.27-4.57 (m, 7H), 5.10 (s, 2H), 6.63
(s, 1H), 7.10 (s, 1H), 7.23-7.60 (m, 5H).
Elemental analysis (C ₁₉ H ₂₂ N ₂ O ₅ S) C, H, N.

Step c 5-(benzyloxy)-1,2,3,
6,7,8-hexahydro-3-(methylsulfonyl)-8-(methylthio)-7
-Oxobenzo[1,2-b:4,3-
b′] Dipyrrole-1-methanol acetate ethyl (methylthio) acetate (5.2 ml,
40 mmol) and sulfuryl chloride (3.2 ml, 40 mmol) were added to dry methylene chloride under argon at -78°C and the mixture was stirred for 20 minutes. to this
Step b compound (15 g, 38 mmol) over 2.5 hours
and 1,8-bis(diethylmethylamino)naphthalene (proton sponge, 7.9 g, 37 mmol)
A solution of 100% of the solution in dry methylene chloride (250ml) was added.
After another 1.6 hours, triethylamine (5.5 ml,
39 mmol) was added. After 30 minutes, the mixture was warmed to room temperature. This was diluted with methylene chloride, washed with water, dried (Na ₂ SO ₄ ) and concentrated. (Methylthio) of the Sommelet-Hauser product with glacial acetic acid (57 ml)
Ring closure to oxindole was carried out for 1 hour at room temperature. After evaporation of the acetic acid, a residue remained and was taken up in ethyl acetate and washed with 5% KHSO ₄ , 5% NaHCO ₃ and saturated NaCl (in some experiments, most of the oxindole precipitated out of solution during work-up). and collected by filtration). The organic layer was dried (Na ₂ SO ₄ ), concentrated and chromatographed on silica gel (ethyl acetate, hexanes).
The title compound was obtained (14.1 g, 77%). NMR
(CDCl ₃ ), δ2.02 (s, 3H), 2.06 (s, 3H), 2.80
(s, 3H), 3.63-4.57 (m, 6H), 5.18 (s, 2H),
7.20 (s, 1H), 7.20-7.61 (m, 5H), 8.06-8.30
(br.s, 1H); Calculated value for MS C ₂₂ H ₂₄ N ₂ O ₆ S ₂
476.1076, actual value 476.1063, m/e430, 337,
289, 199.

Step d 5-(benzyloxy)-1,2,3,6,
7,8-hexahydro-8-methyl-3
-(methylsulfonyl)-8-(methylthio)-7-oxobenzo[1,2-b:
4,3-b']dipyrrole-1-methanol acetate.

Compound of step c (10.2 g, 21 mmol) and Na ₂ SO ₃ in acetone (16 ml) and DMF (162 ml)
To a vigorously stirred mixture under argon of (36 g, 0.34 mol) was added methyl iodide (12 ml, 0.19
mole). After 4.5 hours, the mixture was concentrated and the residue was dissolved in methylene chloride and water. Organic layer with 1NHCl
and saturated NaCl, dried (Na ₂ SO ₄ ), concentrated and chromatographed on silica gel. Elution with acetone in methylene chloride gave the diastereomeric product of the title compound as a pink solid (8.8 g, 84% yield): 1R (neat) 3150, 2970, 1700, 1605, 1440 ,1225,
1150cm ^-1 ; NMR (CDCl ₃ ), δ1.74, small amount of diacetreomer, 1.81, large amount of (s, 3H), 1.92, large amount of diastereomer, 1.95, small amount (s, 3H),
2.10 (s, 3H), 2.84 (s, 3H), 3.75−4.45 (m,
5H), 5.18 (s, 2H), 7.22 (s, 1H), 7.5 (m,
5H), 8.26 (br s, 1H).

Step e 3-methyl-1-oxa-4-thian-
2-on.

Ethyl 2-bromopropionate (50g,
0.276 mol) and 2-mercabutoethanol (21 ml,
0.30 mol) was refluxed with crushed anhydrous K ₂ CO ₃ (42.0 g, 0.304 mol) in 0.31 g of acetone under nitrogen for 18 hours. The mixture was cooled, filtered and the solids were washed with acetone. The filtrate was concentrated to give 50 g of colorless oil. To this oil dissolved in 230 ml of 9:1 methanol/water was added dropwise KOH (17.0 g, 0.303 mol) dissolved in 170 ml of 9:1 methanol/water and the mixture was stirred for 2 hours. This was cooled to 0-5° C. and neutralized with 52.0 ml (0.315 mol) of 50% (vol/vol) HCl. After 30 minutes at ambient temperature, the solvent was removed in vacuo and the residue was dissolved in 0.41 toluene. The solution was refluxed for 2 hours with azeotropic removal of water produced during the reaction. The mixture was then cooled, then concentrated and chromatographed on silica gel, eluting with 25% ethyl acetate in hexane.
24 g of colorless oil was obtained. This was distilled bulb-to-bulb at 0.35mmHg and 80-85℃, yielding 22g (60%).
The title compound was obtained as a white solid. Melting point 44-45
℃; 1R (nujiol) 1739, 1725, 1454, 1379,
1296, 1238, 1202, 1162, 1151 cm ^-1 ; NMR
(CDCl ₃ ), δ1.45 (d, J=7Hz, 3H), 2.9−3.4
(m, 2H), 3.85 (q, J=7Hz, 1H), 4.3−4.8
(m, 2H), analysis (C ₅ H ₈ O ₂ S) C, H, S.

Step f 5-(benzyloxy)-1,2,3,6,
7,8-hexahydro-8-methyl-3
-(Methylsulfonyl)-8-[(2-hydroxyethyl)thio]-7-oxysobenzo[1,2-b:4,3-b']dipyrrole-1-methanol, acetate.

Sulfuryl chloride (17.8 ml, 0.22 mol) at -70℃
to a solution of the compound of step e (30.0 g, 1.227 mol) in dry methylene chloride under nitrogen,
The mixture was stirred at this temperature for 45 minutes. A solution of the compound of step b (83.9 g, 0.215 mol) and the proton sponge (47.1 g, 0.220 mol) in 1.51 methylene chloride was added to the mixture over 2 hours while maintaining the reaction temperature at -70°C. 2 more red suspension
The mixture was stirred for an hour and then triethylamine (31.6 ml, 0.227 mol) in 100 ml of methylene chloride was added dropwise over 15 minutes. The mixture was warmed from -70°C to 0°C over 2 hours and the mixture turned black with edges.
The reaction mixture was poured into 4L of saturated NaCl. The aqueous layer was extracted with methylene chloride and the combined organic layers were concentrated and chromatographed on silica gel. Elution with acetone in methylene chloride and trituration of the collected product with acetone yields
57.5 g (51%) of the title compound (mixture of diastereomers) was obtained as a brown solid. melting point
184-204℃; 1R (Nujiyoru) 3165, 1740,
1710, 1635, 1490, 1460, 1455, 1445, 1345,
1335, 1230, 1220, 1160, ^{1155, 1030cm -1} :
NMR (CDCl ₃ ), δ1.6−1.9 (m, 4H), 2.1 (s,
3H), 2.3-2.8 (m, 3H), 2.8 (s, 3H), 3.4-4.3
(m, 6H), 5.2 (s, 2H), 7.2 (s, 1H), 7.3−
7.6 (m, 5H), 8.3 (br s, 1H), analysis (C ₂₄ H ₂₈ O ₇
N ₂ S) C, H, N, S.

Step g 5-(benzyloxy)-1,2,3,6-
Tetrahydro-8-methyl-3-(methylsulfonyl)benzo[1,2-b:
4,3-b′]dipyrrole-1-methanol borane-methyl sulfide (2.0M in THF,
275 ml, 0.55 mol) over 45 minutes to produce 25 g of 4-
Compound of stage f (57.5) in dry THF (1.21) containing molecular sieves and kept under nitrogen
g, 0.11 mol). Note! Vigorous foaming and gas evolution occurs. The solution was refluxed for 3 hours,
Cool to 0° C. and stop by carefully adding 0.51 1M HCl dropwise. Note! There is gas generation. The mixture is stirred for 1 hour at 5°C, then diluted with ethyl acetate. The aqueous layer was extracted with more ethyl acetate, the combined organic layers were washed with saturated NaCl,
Dry (Na ₂ SO ₄ ) and concentrate. Chromatography on silica gel eluting with 5% acetone in methylene chloride gave 38.6 g (90%) of the title compound. Melting point 143-150℃. lR (nujiyor) 3380, 3265, 1590, 1465, 1455, 1440,
1330, 1320, 1300, 1185, 1155, 1115, 1100,
1090, 1025, 735 cm ^-1 ; NMR (CDCl ₃ ), δ1.6 (s,
1H), 2.37 (s, 3H), 2.75 (s, 3H), 3.5-4.3
(m, 5H), 5.2 (s, 2H), 7.0 (br s, 1H), 7.13
(s, 1H), 7.2-7.4 (m, 5H), 8.3 (br s,
1H); UV (ethanol) 211 (ε25900), 235
(40000), 269 (5500), 306nm (5500); MS C ₂₀
Calculated m/e386.1300 for H ₂₂ N ₂ O ₄ S, measured value
386.1315. Analysis ( _C20H22N2O4S )C _{, H} , _N .

Reference example 7 1,2,8,8a-cyclopropa[c]
Preparation of benzo[1,2-b:4,3-b']dipyrrol-4(5H)-one (12), N-(methylsulfonyl)-4,5-pyrrolo-6-hydroxyindoline bromide (step 10 ), but it is expensive
If instead of isolating the product bromide at Rf (0.64), the reaction mixture before work-up is concentrated in vacuo and applied directly to a thick layer of silica gel plate.
A new low Rf band (0.32) was observed, compound (12)
will be collected as. NMR (CDCl ₃ ): 9.5 (br.s, 1H), 6.83 (dd, H _a ),
6.34 (s, H _b ), 4.10 (d, H _c ),
3.93 (dd, H _d ), 3.04 (s, 3H),
2.93 (m, H _e ), 2.00 (d, 3H),
1.97 (dd, H _f ), 1.37 (dd, H _g ).

J _c,e =0.0Hz J _e,g =4.4 J _c,d =9.7 J _f,g =4.4 J _d,e =4.7 J _NH,a =2.0 J _e,f =7.7 J _a , CH ₃ =
<1.0 MS: Silylation with BSTFA (DMF containing 1% TMS-Cl) is m/
yielding eM ⁺ 386/388 (22, 12% corresponding to product + Me ₃ SiCl).

UV: (methanol) λ224, 272, 338 This compound was tested using a standard test tube dilution assay.
The culture medium was assayed against L1210 Mus musculus leukemia cells, and the following results were obtained.

ID ₅₀ (μg/ml) = 0.13 ID ₉₀ (μg/ml) =
0.42 Reference Example 8 Alternative Preparation of Compound (12) 0.1 mmol of diisopropylethylamine was added to 4,5-pyrrolo-6-hydroxyindoline bromide (or mesylate) (0.1 mmol) in 1 ml of methylene chloride and the mixture was heated under N ₂ at room temperature. Stir for 24 hours. The reaction solution is taken up in 10 ml of methylene chloride, washed with 0.1N HCl, dried over Na ₂ SO ₄ and concentrated to yield the desired product. Further purification can be achieved by silica gel chromatography.

Compounds (11) and (12) exhibit antibacterial activity against Bacillus subtilis, Klebsiella pneumoniae, S. lutea, Staphylococcus aureus, and M. avium.

Definition R ₁ = CH ₃ −, −CH ₂ Ph, CH ₂ = CHCH ₂ −, −CH ₂
SCH ₃ , −CH ₂ OCH ₃ , −CH ₂ OCH ₂ CH ₂
OCH ₃ , -CH ₂ CCl ₃ , -CH ₂ CH ₂ Si(R ₂ ) ₃ R ₂ , R' ₂ = alkyl (C ₁ - _{C 5} ), phenyl, hydrogen, R ₃ = o-alkyl (C ₁ - C ₂ ), alkyl (C ₁ ~
C ₅ ), phenyl [Note: R ₃ is only o-alkyl for compound (2)] R ₄ = SO ₂ R ₂ , SO ₂ CH ₂ COPh, CO ₂ CH ₂ Z X = SO ₂ R ₂ , Cl , Br, I Y=Li, Na, K, MgX Z= _CH2I , _CCl3 , _{CH2SO2R2 ,} Ph, _{fluorenylmethyl} 1 to 2 or 1 to 5 used herein Alkyl carbon atoms include methyl, ethyl, propyl,
Includes butyl, pentyl, and their branched chain isomers.

Claims (1)

[Scope of Claims] 1 A compound of the formula [Formula] [wherein R ₁ is selected from the group consisting of H, CH ₃ −, and -CH ₂ Ph, and R ₂ is an alkyl of 1 to 5 carbon atoms] Yes, R ₄ is SO ₂ R ₂ and X is OSO ₂
selected from the group consisting of R ₂ , Cl, Br, and OH]. 2 A compound of the formula [Formula] [wherein R ₂ is alkyl of 1 to 5 carbon atoms and R ₄ is SO ₂ R ₂ ] can be converted into alkanesulfonyl chloride, carbon tetrachloride/triphenylphosphine, and carbon tetrabromide/triphenylphosphine, wherein R ₂ and R ₄ are as defined above,
X is selected from the group consisting _{of -OSO2R2} , Cl, and Br. Claim 2 consists of reacting a compound of formula 3 with triphenylphosphine/carbon tetrabromide.
A method for producing a compound of formula [formula] according to the following.