JPH0314561A - Antibiotic cc-1065 fragment - Google Patents

Abstract

NEW MATERIAL: A compound of formula 9 [wherein R₁ is H, CH₃, CH₂Ph, CH₂CH=CH₂, CH₂SCH₃, CH₂OCH₃, etc.; R₂, R₃ are H, a 1-5C alkyl, phenyl; R₄ is SO₂R₂. SO₂CH₂CO phenyl, CO₂CH₂Z (wherein Z is CH₂I, CCl₃, phenyl, etc.), Y is NH₂, NO₂, H; X is OSO₂R₂, Cl, Br, I, OCOCH₃].

USE: Active against certain kinds of bacteria, e.g. Bacillus subtilis, Klebsiella pneumonia, Staphylococcus aureus, etc., and used for disinfecting a pile of washed dishes or as an antibacterial rinsing agent for laundry.

PROCESS: A compound of formula 2, obtained through the reaction of a malonate and a β-ketoester of a compound of formula 1, is reduced to obtain a compound of formula 3. Then, the compound of the formula 3, after its functional group being exchanged, is reduced and cyclized to obtain a compound of formula 5 (which is the compound of formula 1 in which Y is H). Furthermore, the unstable groups are substituted, nitrated or reduced to obtain the corresponding compounds of formula 6 to formula 8.

COPYRIGHT: (C)1991,JPO

Description

DETAILED DESCRIPTION OF THE INVENTION Antibiotic CC-1065 is disclosed in U.S. Pat.
No. 888, disclosed and claimed in terms of chemical and physical variables. Subsequently, the structure of antibiotic CC-1065 was published as "Structure of Antibiotic CC-10650".
tureProof of Amtibio'tic
cc-1065) J TG March 7
(D, G, Martin), Chidestar (CG, Chidestar), T-';
．． J. Duchamp), and S.A. Mizsak, J. Antibio
It was clarified and elucidated in vol. 33, p. 902 (1980).

The structure of antibiotic CC-1065 is shown in Formula 1. The antibiotic CC-1065 consists of a system of three fragments, the most unstable part of the molecule being 1, 2, 8, 8 & - cyclopropyl
C]benzoC 1.2-'b :4,3-b')dipyrrole-4(5H) monoone, which is designated herein as compound 0z. Attempts to obtain this fragment by antibiotic CC-10650 reduction were unsuccessful. For this reason, it is created using the floor method. The present compound, as well as certain intermediates disclosed in the unknown specification, may be isolated from certain bacteria, such as Bacillus subtilis.
), Sarc.
ina lutea), Staphylococcus aureus, and Myco/
Z Cuterium avium M (Mycobactsri
um avium) Active towards virtue. 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. 4,169,8815. 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 t12 is shown in 2. This stage is as follows.

Step 1: First step of combination or clue (nucleophilic substitution of aromatic)
is G. Gourdais (.T, Bou-rcrai
S) and C.Mahieu, Co.
mpt, Redux [C], vol. 263, p. 84 (19
1966). Also, G. Bourdais and C. Germain (C, Germain), T.
See also, et. Letters 195 (1970). The various R groups can be modified according to procedures described in the literature (appropriate references detailed under step 8) (1
can be introduced on top of the phenol precursor. The various malonates, β-ketoesters and β-diketones used are all known compounds.

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

Step 3 Monofunctional group exchange. In particular, the chemistry described herein is
X=! This is the case of 302CH3. 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. Halogen analogs of 4 can be obtained by standard procedures known in the art such as Ph3PICC (CBr4) and N-:i-dosuccinimide/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 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 alternative reducing situations, Fe or Ticz3 or sncz2 in acid
can be used. This would then require another step involving base treatment to cause cyclization to (5). An example of reduction with iron is Fe/CH3CO2H/CH
Use 3CH20Ht (u-・S・Ponticello (G,
S., Fonticello) and J.J.
Baldwy 7 (1, I, BaMwin)
,・J, Org, Chem, Volume 44, Page 4003 (
(1979) ). R, =CH2Ph or -CH2C
These conditions are necessary when H=CH2.

The concept of nitro reduction followed by in situ cyclization to indole was proposed by A.D. Batch-i (A,
D, Eatcho) and G Stage 5 Brew Lime Grew 7S - (W. Lein
-Gruber), German Published Patent Application No. 20578
No. 40 (1971), which was advanced by the old Reias reductive cyclization to indole.
ert) procedure (R.J. Sundberg).
), “The Chemist”
ry of engineering and leg) J pages 176-183, 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 into acetate or C1-C5 under standard conditions (alkali carboxylate in acetone, DMF or alcohol).
(al-Jl) with the conjugate base of the carboxylic acid. Since some hydrolysis may also occur when XzSO2CH3, the reaction mixture is treated with acetic anhydride before isolation of (6).

Step 6 - Dioxidation is carried out using acetic acid, acetic anhydride, sulfuric acid,
It can be carried out under various conditions described in the literature including R/H20, 7 alcohol and nitric acid in hinitroalkanes. The regioselectivity of this reaction is supported by the spectroscopic data obtained.

Stage fV7. --Reduction of the 4-unit to the ami-7 group follows the same chemical reaction description in step 4, but omitting the base.

Step 8 - Indole combination. This procedure is outlined in Summannoindole Chemistry [H.G.
Assman) et al., J. Am, Chem, Sac
．． g6 volume 5494 pages, 5508 pages, 55l2 pages (19
Based on 1974]. However, it requires some modifications not disclosed or suggested in Suman's study. The chemical sequence and some intermediates are shown in Equation 3.

α-thiomethyl esters are known. This method uses chlorosulfonium complex A and reacts it with aniline (6), which deviates from the published but Suman route. Gasuman created a black amine of aniline and reacted it with a thioether to produce oxindole. Second, the method uses two different bases, whereas Gassmann uses Anili 72 equivalents followed by Base 2. Triethylamine, diisopropylethylamine, bis(1.8l) Preferable is bis(l,
8-dimethylamine) naphthalene, with triethylamine as base 2. A variety of solvents can be used, such as chloroform, acetonitrile, tetrahydrofuran (THF') and methylene arsenide chloride, the latter being preferred. The temperature range is from one edge to -80', and the reaction is carried out under an inert atmosphere. Cyclization to oxindole B is
As described by Gassmann, acid catalysis (2
Best facilitated by N HCL, Step 9 ether and/or ethyl acetate).

The final reduction (reductive removal) to (9) can be achieved with lithium aluminum hydride (Tokai PJ as described by Gassman) or dibora/type reagents, the latter being by far superior.

The preferred reagent is (C!) in THF for 24 hours at room temperature.
{3), S. It is BH3.

This deprotection step (removal of Rl) was performed using Rl in Example 8.
: Described in detail in CH3.

Many procedures have been described in this technique, including the cleavage of methyl ether, but under an inert atmosphere (95-100”)
hexamethylphosphoric triamide (HMPA)
) were found to be effective (S.C. Welch and A.S.C.P.).
Rao), Tet, Let'ters No. 505 (
1977) and T.R.K.IJ-(
T.R.Kelly), H.M. Dali (H.
M. Dali) and Dapliu Ji Tsang (
W. G, Tsang), Tet, Lett8rs
, No. 3859 (1977), or Me2S-BBr3 in dichloroethane (P.G.
F., G., tVillard) and C.P.
Fryhls (C.B.), Tet, L.
etters No. 3731 (1980)].

When R1=CH,Ph, standard hydrogenolysis conditions (
H2, Pd/C) is well suited for deprotection (O
rg, Reactions+ Vol. 7, p. 263 (19
5: Year)]. When R = CH2SCH3, mercuric chloride in acetonitrile/H20 removes the ether (R.A. Holton
) and 7 - Rou'; - ・7' - Pi, c (
R,G, Davis), Tet. Le'tters
No. 533 (1977)]. R1=CH20CH3
When , a mild acid such as acetic acid produces phenol 10 [J, Mea, Chem, Vol. 9, p. 1 (196
6) or 8ynthesis 244 (1976)
Year)〕. In practice, however, this protecting group is lost in step 6, but can be reintroduced prior to step 7 under standard conditions. R
1: -CH20CH2CH20CH3, 7 enol can be made with ZnBr2 or TiC of CH2Clz'P (Tst. Letters No. 3Q9 (1
976)]. R = -CH2CH: When CH2, several two-step procedures carry out the deprotection of the ether [Pd in alcohol/CAng. Chem, engineering
t, Ka. Volume 15, page 558 (1976); Sea, CH3C02H, Tet, Le in rioxane
tters No. 2885 (1970); t-Bul
l! :, DMSO, I{2So in acetone,
, J, Cham, SOc, 1903 (1965); RhC4 (PPh3)3 in alcohol, DABC
O, followed by pH 2, J, Org, Chem. 3g
Volume 3224 (1973)]. Rlm 1CH2C
When H2S i (R2) is 30, deprotection is performed with Bu, NF
'H.Gerl Rank (H.Gerl)
ach) etc., Helv, Chin, Acta, 5Q
Volume 3039 (1977)].

See Attack Floor 101 Stage 3.

Step 11 - This role (when X=Br) is promoted in contact with silica gel and! It also occurs when left in a protic solvent. This reaction can be carried out using tertiary amines, bases such as pyridine, t-ptokind, etc. ! It also proceeds in the presence of aqueous weak bases such as salts and carbonates.

The following examples are illustrative of the materials and methods of the invention, but should not be considered limiting. Unless otherwise noted, all percentages are tt, melted t! # All mixture proportions are by volume.

Mi 12 Mi Aryl diethyl malonate (2) to 2-
Preparation of aryl-1,3-propanediol (3) THF in N2 cooled in an ice-water bath 400 1ffj
100 t (0.70 mol) of diimptylaluminium hydride (D-BAL) in 400 toluene
Add. Add arylmalone in TIP 100 to this stirred solution +21 33. t) f (
0.105 mol) is added. 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 to cold 3 N HCt (approximately 1.5 2 ) in portions with stirring.

The mixture was then diluted with KtOAc l followed by cH2Ct2
1000ml! f Extract. The combined organic phase is
a2SO. dried on top, leaving a reddish-brown residue (21.:1M
) Concentrate at t. 6 on residue medicine r nore 5007
0% EtOAc/hexane → 10Q qb mtoA
Chromatography with eluent with a gradient of diol +31 (U-62,598) 11.751'
(Yield: 49%) is produced as a pale red oil (it will harden if left standing with frozen honey).

NMR (cncz3): 7.5 ~ 7.0 (
m. 31{), 3.80 (8.3H), 4.
0-3.3 (m, including rH-20H).

MS, C10H, No5 Calculated value: 227.0
794 Measured Value 2 227.0780 Analysis: Calculated Value: C, 52.86, H. 5.76
, N, 6.16 Measured value: C, 53.40. H
．． 5.77. M, 5'. 99 completed! ” 2-aryl 1.3-glopanediol {3} to 2-7 reel-1.3-fo pandiol bismesylene} 14
) of methanesulfonyl chloride 6.89 (tl.
06 mol) is added. 5°? After stirring for 30 minutes at room temperature (a), the solution was concentrated in vacuo and CH2Cl=/
Taken up during l'N HCI. Separate the organic phase and
2So, dry on top, residue! Concentrate with EtOAc
Trituration gave an off-white solid, and the mother liquor was chromatographed on silica gel (K.tOAc eluent), total yield -ml 6. fi5P (yield 86
%), melting point 122-123'' (recrystallized from acetone)
Compound (4) Bismesylate (σ-62,597)
will occur.

NMR (Acet-d5): 7.7-7.2 (
m, 3H), 4.62 (d, 4H, I
= J [Iz), 4.11 ( ”+ I H, J = 7
Hz), 3.92 (8.3H), 3-06 (8,
5H).

Analysis: Calculated value C for C1■HL7No9S2,
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.

IDao (μg/at) = 6. υ Engineering D
9a (ltg/ml) = 18 Example 1 2-
Aryl-1,3-propanediol bismesylate (
4) to 6-methoxyindoline bismesylate (5)
THF 3Q d; ICtOAc, and compound f41 in 150% absolute ethanol 1.915
' (0.005 mol) and add triethylamine L5- and PtO2400. While shaking this solution, the hydrogen pressure was increased to 0.492 - 0.703Ka 2 (7 -
10 psi) Place the blade below. The reaction solution was then filtered over Celite and rl! in vacuo! do. CH several times in vacuum
, CL, and then convert the residue into C Fl2CL2
100 ml and cooled under N2 in an ice bath. 1.5 parts of triethylamine are added to the stirred solution, followed by dropwise addition of 900 μt of methanesulfonyl chloride. After stirring the blade, let the solution come to room temperature for ω minutes! ,\
Make it. Next, wash with solution I N HCt, Na2So,
Dry on top and concentrate. 60% residue on silica 150y! Rapid chromatography with 1:tOAc/hexane 500- followed by 80% eluent 1000- to give an off-white solid, mp 122-123° (
Compound (5) (recrystallized from ethanol), 6-methoxyindoline bismesylate (U-62.586) 1
．． 37 (yield 78) was recovered.

NMR (DMF-dy): 7.36 (d,L
H, I = 8.5 Fiz), 7. 00
(d, IH, J=2FIz), 6.69(da, II{
, J=2. 8.5Hz), 4.47 (2H,
a. J = 6Hz), 4.3-3.6 (m
, 3H), 3.80 (s, 3H)+
3.2Ll. (8.3HC.3.07 (s,
3H).

Analysis: Calculated value C, for C2HX7NS20,
42.97, H, 5.11, N. 4.18 Measured value C, 42.87, H. 5.27, N, 4
．． 29 This compound was assayed against L1210/- murine leukemia cells in culture by a standard in vitro dilution assay with the following results.

IDso (μg/mj) = 4.8 D9o (μg
/1) ='IO Example z Preparation of 6-methoxyindoline bismesylate (5) to 6-methoxyindoline acetate (6) 6-methoxyindoline bismesylate (5) in DMF 3Qd 13.02 (39 800 mmol of absolute ethanol followed by 322 mmol of sodium acetate are added.

The heterogeneous mixture is refluxed under N2 for 24 hours, cooled and condensed in vacuo. Blend the residue with vinegar? ! 100 d
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 Ha2So, filtered through charcoal and concentrated with oil 1. If the oil is solid, it is a compound (6
) of 6-methoxyindoline acetate (100%, further purification possible by silica chromatography using 60% EtOAc/hexanes eluent if necessary).

NMR (CDCz3): 7.17 (d,I
H, J=8.5Hz), 7-02(d, 1”+ J=
2[1z>,6-60 (dd.IJJ=2.8.
5Il[z). 4.18 (d. 2H. J=6
[1z), 4.1-3.4(m, 3EI), 3.78(
8.3H), 2.91 (s, 3H). 2. o5
(s, 3H).

Analysis C13H,. For NO, S, calculated values: C, 52.16, H, 5.76. N
, 4.68 Measured value: C, 52.13, H. 5
．． 79, N, 5.27th day; Calculated value: 299. U
827 Measured value: 299.0823 Example 3 6-methoxyintolyne acetate {6}
of 5-nitro-6-methoxyinto phosphoacetate (7)! 500 mg (1.67 mmol) of 6-methoxyinto-phosphoric acetate f6) in 20 mt of ditromethane from Il
Add 90 μt of HNOs to the solution. cooled (O
-d) Separate the reaction solution and dry over Na, So,
Concentrate. The residue is silica 50? Upper chromatography (fiO%!!itOAc/hexane eluent → 1
When applied to (10 rumored KtOAC), it becomes a yellow solid with a melting point of 1.
75-177° (recrystallized from ethanol) Compound (7
), 5-2}-6-methoxyindoline acetate (
σ−62.696 ) 440! ! lI (yield 76%
).

Njuk (DMF-d?): 7.91 (g, I
H). 7.20 (g, IH). 4.27
(d, 2J J=611z), 4.3-3.7(m
, 3FK), 3.98 (a.3H), 3.17 (+3,
3H), 2.07 (8, 3H).

Ikuchika: C-e, H-e. Calculated value for N, Owas C, 45.34, H, 4.68, N,
8.14 Measured value C, 44.81, H, 4.7
7. N, 8.16 This compound was assayed against L121t) Mus musculus leukemia cells in culture in a standard in vitro dilution assay with the following results.

ID5o(μg/d)=)so 工D*o(μg/J
=>F4 real'llll+ iFl of 5-nitro-6-methoxyindoline acetate (7) to 5-amino-6-methoxyindoline acetate (8)! THIP5Q
To 4.52 (13 mmol) of 5-nitro-6-methoxyindoline acetate (7) in 150 ml of absolute and absolute ethanol was added 500 η of Pt02, and 500 η of Pt02 was added.
Intake stops below 703Kf/α” (10psi)1
Shake for about ω minutes. Filtered and concentrated in vacuo.

After concentration, the product 3. Of is precipitated. This is filtered off and the mother liquor is quickly chromatographed with KtOAC solution over silica Glue 1002, yielding an additional 0.62.

Total yield (3.6) of 5'-amino-6-methoxyindoline acetate (σ-62.697) of compound (8)
9') has a melting point of 134-135°, (from acetone/cyclohexane).

Lu {P. (CDCt3): 7.02 (11.
IH), 6.65 (8, IH), 4.16 (d, 2H
, J=6Hz), 4.1-3.5(m, 3H). 3.8
3 (8.3H). 3.6 (br.s, 2H), 2.83
(s+3H).

Analysis: Calculation platform C for CL3H18'205S,
49.67, H. 5.77, N, L91 measurement C, 49.74, ” + 5.72, H, 8.94 This compound was tested in a standard test tube dilution assay by diluting 1 of the culture medium.
The assay was performed on 1210 Mus musculus leukemia cells and the following results were obtained.

Dso (μg/ml) =>50 Dso
(μg/1)=>5049Pj@13 5-amino-6-methoxyindoline acetate (8) to 4,5-
mW of Pishoro 6-methoxyindoline (9) Cld”b to CH2Ct214- dried under nitrogen at -7
"Lt solution (20μt Ctd ml CH2CL2
) Add 6-0-. Add cH3 to this stirred solution.
(CH,S)CHCO2C2H537gμt (2.5
mmol) [E-H-Wink (]!:, Fi,
Tick), Tea Ya? Nishi (T, Yal!l
anilihi), x7f・shi-wa-sai? − (
H. C., Wertheimer), Y.E.
Hof (Y, E. HQf!), BP Proctor and S, A. Goldblith, J.
Agr, Food Chem, Volume 9, Page 289 (
(1961), CH3(Br)CHCO2
prepared from C2H5 and methylmercadetide]. 5
After minutes, dry CH2Ct2 3. 470 1 nq (2.2 mmol) of 1,8-pisdimethylaminonaphthalene in U1 and 628 nq (2.2 mmol) of AniIJ/Intolin f8)
．． 0 mmol) solution is added dropwise over 15 minutes. Stir the red solution at -75° for 2 hours, then CH2CL2
35 υ μt of triethylamine in 650 μt are added dropwise over a few minutes. Remove cooling bath.

When the reaction solution reaches room temperature, it is briefly concentrated in vacuo. Add 5 d of KtOAC, 5 d of So-Tel, and 2 N HCt 6 +d to the residue and stir vigorously for 2 hours. Separate the organic phase and dilute the aqueous layer with KtOAc/Kt
Extract at a ratio of 20 (1:1). Combine the organic phases and add N
Dry over a2So, and concentrate. At this point, the residue was taken up in 10 ml of THF and diluted with 2M BH3.SMe2.
3-0- overnight at room temperature under nitrogen. Alternatively, the diastereomeric oxindole CBI derived from acid treatment can be isolated at this point by silica chromatography (507, (7)% KtOAC/hexane to (7)% KtOAc/hexane). .

GC-MS: m/e M+414 (15%),
227 (100%) -2'1<81! :-3
9 NMR (CDC/s): 8.4 (br, 8
, LH). 7.11 (8+ IH). 4.5-3.7
(m, 5H). 3.90 (8,3H). 2.95 (
8,3H). 2.10 (8,3H). 1.92 (
a. 3H), 1.82 (8.3H) - Major diastereomer (2.5/1): Minor diastereomer shows the following differences: -1.99 for SCH3 and -CH3 (
θ, 3a), x. 76? θ, 3H), NH is 7.7.
The CH2 band is at 4.3-3.6 ppm.

The aqueous phase from the acid treatment is neutralized and extracted with CH2Ct.

The C H2C L solution was dried, concentrated, and chromatographed on silica gel with 50% acetone/cyclohexane to recover starting material (anilinoindoline) and dearylated starting material. yields 20%.

The 0-reducing removal reaction of borane dimethyl sulfite (to B) stops gas generation! It was then stopped and finished by NHct, and taken up during CH2Cz2/H20.

The separated organic phase is dried over Ha2So and concentrated.

The residue was chromatographed on silica gel (5f')%
acetone/cyclohexane) to produce the product (9H
55η. Minor yield 75%, 85% based on recovered starting material. Melting point 182~18f (160″
phase change, recrystallized from chloroform). Product 4,5-
Pyrrolo 6-methoxyindoline (9) CU-62
, 233).

NMR (CDCt3): 8,3 (br,
B, IH), 6.96 (8, 2H), 4 -
2-3. 5 (m, 5 H + OH) +
3 - 9 2 (s + 3H), 2.87 (8, 3
EI), 2.41 (8.3H).

Analysis For 014”18N204s, calculated value C,
54.17. H, 5.84, N, 9. o3 measurement value C, 53.49. H. 5.96, N, 9
．． 42GC-MS: O-A7T} - m/eM
+352 (13%), 213 (many 00) -2'-
1 multi-SK-blade, concentration 150-260° (107 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.

Engineering D5o (μg/+++/) =>4 I”%o (
μg/+d) =>408 Example 4 - 4.5 - Preparation of 4,5-pyrrolo-6-hydroxyindoline 0I from hyrolow-6-methoxyindoline (9) Dry and degassed HMPA (hexamethylphosphorus triamide) 10 - to butyl mercabutane 35 at room temperature
Add 0μt. Cool the solution in an ice-water bath and add 1.5 M n-BuLi 2,0- in hexane dropwise. The reaction was cooled down to room temperature, then indole (9) 1'OO
Add WIIi (0.3 mmol) with stirring. Heat the solution to 100° for 2.5 hours. TLC (5Q
% acetone/cyclohexane);
Heating is terminated when the conversion is approximately 75° complete (by phosphorus/a acid spray). The cooled solution was added to IN HC.
Pour into t(1oo*) and add 20 ml +! :Extract with tOAc. Wash the separated organic phase with an additional 50 g of water.

The aqueous phases are combined and back extracted with EtOAC 20d. The organic phases were then combined and washed with NazSo. dry on top, concentrate in vacuo and place on a silica gel 10070 column to 50%
Elute with a7ton/cyclohexane. Starting material 25η
and 4,5-pyrrolo-6-hydroxyintoline (ff-62,370) of raw αO (isolated yield of 44 excellent, 69% based on recovered starting material)
.

NMR (Acet-d6): 7.8 (br,
s, IH), 7.03 (θ, 1u), 6.8
3 (8. IH). 4.25 -3.25
(m.5H), 2.86 (11.3K),
2.36 (S,3H) This product was dissolved in acetic anhydride 1. Or! Lt and NaOAC20
Treat at room temperature overnight and pick up at CH, Ct, /H20゜. The organic phase is separated, dried over Ha, So, and concentrated.

NMR (CDCz3): 7.8 (br.B.
IH), 7.16 (8, IH), 6.97 (B.
IH). 4.42. 4.20 (d(L,
2H), 4.2-3.7(m, 3H). 2.86
(i1, 3H). 2.40 (8,3H), 2.35 (s
,3H),2.05(s,3I{)GC-MS: m
/eM"38u (254), 199 (10o'
l) -2/-14SE This compound was assayed against L 1210... murine leukemia cells in culture by a standard in vitro dilution assay with the following results.

ID,. (μg/J=>5 engineering D,. (μg/+d)
=>5fAf+l S 4.5 - Hirolo-6-human 9-loxyinto-9-phosphorus alcohol (4,s from 1G
- Preparation of Hirolow 6-hydroxyindoline bromide Uυ alcohol substrate in 1.0 ml of dry acetonitrile
rMi (65 micromoles) was treated with CBr,
33■ (100 μmol) and human rifhenylphosphine (ph3p) 26η (100 μmol) are added. After additional stirring, additional CBr, 11η and P
Add h3P3η. After a total of ω minutes, the reaction was changed to CH2CL2/
It will be taken up during H,O. Separate the organic phase and add Na2So,
Dry on top and shrink. 20 x 20 (250 m
Place the residue on 3 μ silica glue plates and elute with 50% acetone/cyclohexane, product with high 3Rf value (0.64; l'jf of alcohol = 0.45)
, that is, the compound (1υ of 4,5-pyrrolo-6 human 9 loxindoly/bromide (σ-62,694) about 8
Collect ■.

NMR (CDCz3): 8.5 (br. a,
IH), 7.1 CB, IH) 6.9 (El,
IH), 4.23 (a, 2H, J==6Fl
z), 4.0 3.5 (m.3H), 2J9 (B.3
H), 2.38 (8,3H).

Peilstein test: Positive MS: Calculated value 430.0382 Measured value 430.0375 (%
/-TMS): m/eM+430/432 (1
4%), 271 (90%), 147 (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.

XD5a CAFE /td) = 0. 12
D90 (μg/1) = 0.37 D90 4
, 5-pyrrolo-6-hydroxyinto-9-phosphorus alcohol substrate preparation of 4.5-pyrrolo-6-hydroxyindoline mesylate uυ from pyridine (if) 20μ
65 micrommol), do not let it cool down in a water bath.
Add 8 μt (To micromoles) of methanesulfonyl chloride to the bottom. After a minute, add an additional 2 μt of CH, So2CL, and after a total reaction time of ω minutes, finish with 2 N HCz/CH2Cz2.

Separation TLC (20 x 20cm. 250
The high Rf material (3 μ silical plates) (NMR showed only one CH3S02 group. 695) produces 9■.

NMR (Acet-aa): 8.6 (br.
s, IH), 6.97 (Japanese, IH), 6.74 (
fil, IH), 4.3 (m, 2H). 4.1
-3.6 (m, 3H), 2.96 (
fl. 3H). 2.79 (i1.3Et),
2.26 (8, 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.

ID,. (μg/mg) = 1-0 D9Q (μq/
ml')=3.3#A Example 7 1+2+L8a-cyclopro7ξ[C]henso[: 1,z-b: 4.3-
b'] Preparation of Dipyrrole-4(5H) One (1, N-(Methylsulfonyl) One) 4. Following the procedure for making 5-hyrolo-6-hydroquine indoline bromide (Step 1O), but with the hole Rf ( Q.
Instead of using the raw bromide of 64), the reaction mixture before work-up is concentrated in vacuo and a thin layer of silica is added.
When applied directly to the plate, a new lower Rf band (
0.32) was observed and recovered as Compound 02.

NMIR (CDCt3): 9.5 (br.s
, IH), 6.83'(d+1.}k),
6.34(s, H1)), 4.10(d, Ha)+3.
93 (dd, FIdL 3.04 (8,3”
), 2.93 (m.He). 2.00 (d, 3H), 1
．． 97 (da+Hr), 1-37 (a
a,%).

Jc,e=U. OFfz Je,=4・4J(2
, d=9. 7 Jf,g=4・4Jd,
e=4.7 JNR, a=2-OJe,t
= 7.7 Ja, CH, = ( 1.0
MS: BSTFA (1% TMS-Ct
DMF containing )K j silylation is m/eM+3
86 / 388 (22, product + Me, SiCt
12%).

TJv: (methanol) λ 224, 272, 338 This compound was tested for T-
The assay was performed on 1210 Mus musculus leukemia cells and the following results were obtained.

ID50 (Ag/m/) = 0. 13 D90 (μg/nst) = 0. 4 2 calyx example? Preparation of substitute for compound 07J 1 ml of methylene chloride
Add 0.1 mmol of diimpropylethylamine to 4,5-virol-6-hydroxyindoline bromide (or mesylate) (0.1 mmol) in
Stir in a bowl at room temperature for 24 hours. The reaction solution was taken up in l〇l of methylene chloride, washed with υ,l N HCt, and diluted with N
Drying over a2So, and concentration yields the desired product. Further PII can be produced using silica rlucomatography.

Compounds qυ and alpha are derived from Bacillus subtilis, Pneumoniae, S. Lutea, Staphylococcus aureus and M. Shows antibacterial activity against M. avium.

Stage 1 ↓ Stage 2 ↓ Stage 4 (6) ↓ Stage 6 αυ ↓ Stage l1 Only one definition, = CH3-, one CH2Ph, CH2==CHC
H2-, -CH2SCI{. 3, -CH20CH3, -
CH20CH2CH20CH3, -CH2CCI3, (
9} ↓Stage 9? , ), phenyl [Note: R3 is 0 for compound (2)
-Alkyl only] R, == So2R2, So■CEI2COPh,
Co2CH2ZX == So■%, C1, Br, Y = Li, Na, K, MgXZ == CH
2 engineering, CCLI, CH2So2R2, Fh17 #
Olenylmethyl As used herein, alkyl of 1 to 2 or 1 to 5 carbon atoms includes methyl, ethyl, glopyl, butyl, pentyl, and branched chain isomers thereof.

W 4 Emperor A2± {2} (3) U-62,598 room temperature 4 bases 2 (3) (4) U-62,597 Example D (4) (5) U-62,586 Examples (5) (6) Example 3 (6) Example 4 (7) e white example (8) (9》 Keiroku example 5 (7) U-62,696 01 (8) U-62,697 (11 PLf5 example 7 (9) U-62. 23 3 αυ αO U-92,370 {lυ U-62,694 wholesale σ-62,695 11 of U-62,736

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), R_2 and R_3
are hydrogen, alkyl of 1 to 5 carbon atoms, and phenyl, and R_4 is SO_2R_2, SO_2CH_2C
selected from the group consisting of O phenyl, CO_2CH_2Z, where Z is CH_2l, CCl_3, CH_2SO_2R_
2, phenyl and fluorenylmethyl, Y is H_2N-, O_2N- or H, and X is -OSO_2R_2, Cl, Br, I, and -OC
selected from the group consisting of OCH_3].