KR810001407B1 - Preparation of 3-[(S) -1′-phenylethyl amino] propyl amino bromycin - Google Patents
Preparation of 3-[(S) -1′-phenylethyl amino] propyl amino bromycin Download PDFInfo
- Publication number
- KR810001407B1 KR810001407B1 KR7801991A KR780001991A KR810001407B1 KR 810001407 B1 KR810001407 B1 KR 810001407B1 KR 7801991 A KR7801991 A KR 7801991A KR 780001991 A KR780001991 A KR 780001991A KR 810001407 B1 KR810001407 B1 KR 810001407B1
- Authority
- KR
- South Korea
- Prior art keywords
- copper
- acid
- breomycin
- solution
- dissolved
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
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- 125000006308 propyl amino group Chemical group 0.000 title description 6
- 238000002360 preparation method Methods 0.000 title description 4
- 239000002253 acid Substances 0.000 claims description 56
- 150000003839 salts Chemical class 0.000 claims description 7
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 6
- NQONXAAXIFYYMX-JTQLQIEISA-N n'-[(1s)-1-phenylethyl]propane-1,3-diamine Chemical compound NCCCN[C@@H](C)C1=CC=CC=C1 NQONXAAXIFYYMX-JTQLQIEISA-N 0.000 claims description 6
- 231100000252 nontoxic Toxicity 0.000 claims description 6
- 230000003000 nontoxic effect Effects 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 239000002904 solvent Substances 0.000 claims description 2
- ZHHIHQFAUZZMTG-BSVJBJGJSA-N [(2r,3s,4s,5r,6r)-2-[(2r,3s,4s,5s,6s)-2-[(1r,2s)-2-[[6-amino-2-[(1s)-3-amino-1-[[(2s)-2,3-diamino-3-oxopropyl]amino]-3-oxopropyl]-5-methylpyrimidine-4-carbonyl]amino]-3-[[(2r,3s,4s)-3-hydroxy-5-[[(2s,3r)-3-hydroxy-1-oxo-1-[2-[4-[4-[3-[[(1s)-1-phenylethyl] Chemical compound OS(O)(=O)=O.N([C@H](C(=O)N[C@H](C)[C@@H](O)[C@H](C)C(=O)N[C@@H]([C@H](O)C)C(=O)NCCC=1SC=C(N=1)C=1SC=C(N=1)C(=O)NCCCN[C@@H](C)C=1C=CC=CC=1)[C@@H](O[C@H]1[C@H]([C@@H](O)[C@H](O)[C@H](CO)O1)O[C@@H]1[C@H]([C@@H](OC(N)=O)[C@H](O)[C@@H](CO)O1)O)C=1NC=NC=1)C(=O)C1=NC([C@H](CC(N)=O)NC[C@H](N)C(N)=O)=NC(N)=C1C ZHHIHQFAUZZMTG-BSVJBJGJSA-N 0.000 description 79
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 78
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- 229910052802 copper Inorganic materials 0.000 description 66
- 239000010949 copper Substances 0.000 description 66
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 65
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 64
- 239000000243 solution Substances 0.000 description 55
- -1 amino compound Chemical class 0.000 description 51
- 238000000034 method Methods 0.000 description 39
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- 239000003960 organic solvent Substances 0.000 description 4
- VGRXAHXMIDCTNV-UHFFFAOYSA-N (6-chlorobenzotriazol-1-yl) 4-chlorobenzenesulfonate Chemical compound C1=CC(Cl)=CC=C1S(=O)(=O)ON1C2=CC(Cl)=CC=C2N=N1 VGRXAHXMIDCTNV-UHFFFAOYSA-N 0.000 description 3
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 3
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- 206010016654 Fibrosis Diseases 0.000 description 3
- 235000019270 ammonium chloride Nutrition 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
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- 238000005406 washing Methods 0.000 description 3
- RQEUFEKYXDPUSK-ZETCQYMHSA-N (1S)-1-phenylethanamine Chemical compound C[C@H](N)C1=CC=CC=C1 RQEUFEKYXDPUSK-ZETCQYMHSA-N 0.000 description 2
- ADFXKUOMJKEIND-UHFFFAOYSA-N 1,3-dicyclohexylurea Chemical compound C1CCCCC1NC(=O)NC1CCCCC1 ADFXKUOMJKEIND-UHFFFAOYSA-N 0.000 description 2
- DLFVBJFMPXGRIB-UHFFFAOYSA-N Acetamide Chemical compound CC(N)=O DLFVBJFMPXGRIB-UHFFFAOYSA-N 0.000 description 2
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- 241000282472 Canis lupus familiaris Species 0.000 description 2
- 201000009030 Carcinoma Diseases 0.000 description 2
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 2
- JUJWROOIHBZHMG-UHFFFAOYSA-N Pyridine Chemical compound C1=CC=NC=C1 JUJWROOIHBZHMG-UHFFFAOYSA-N 0.000 description 2
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- 238000009825 accumulation Methods 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 125000003277 amino group Chemical group 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 125000004429 atom Chemical group 0.000 description 2
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000012459 cleaning agent Substances 0.000 description 2
- 239000012141 concentrate Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- IXCSERBJSXMMFS-UHFFFAOYSA-N hydrogen chloride Substances Cl.Cl IXCSERBJSXMMFS-UHFFFAOYSA-N 0.000 description 2
- 229910000041 hydrogen chloride Inorganic materials 0.000 description 2
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- 125000002924 primary amino group Chemical group [H]N([H])* 0.000 description 2
- WGYKZJWCGVVSQN-UHFFFAOYSA-N propylamine Chemical group CCCN WGYKZJWCGVVSQN-UHFFFAOYSA-N 0.000 description 2
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- GETQZCLCWQTVFV-UHFFFAOYSA-N trimethylamine Chemical compound CN(C)C GETQZCLCWQTVFV-UHFFFAOYSA-N 0.000 description 2
- RYHBNJHYFVUHQT-UHFFFAOYSA-N 1,4-Dioxane Chemical compound C1COCCO1 RYHBNJHYFVUHQT-UHFFFAOYSA-N 0.000 description 1
- UOFGSWVZMUXXIY-UHFFFAOYSA-N 1,5-Diphenyl-3-thiocarbazone Chemical compound C=1C=CC=CC=1N=NC(=S)NNC1=CC=CC=C1 UOFGSWVZMUXXIY-UHFFFAOYSA-N 0.000 description 1
- 238000001644 13C nuclear magnetic resonance spectroscopy Methods 0.000 description 1
- MRNLLBXPSWMYCK-UHFFFAOYSA-N 2-[2-[2-[[2-[[4-[[2-[[6-amino-2-[3-amino-1-[(2,3-diamino-3-oxopropyl)amino]-3-oxopropyl]-5-methylpyrimidine-4-carbonyl]amino]-3-[3-[4-carbamoyloxy-3,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3-(1h-imidazol-5-y Chemical compound N=1C(C=2SC=C(N=2)C(O)=O)=CSC=1CCNC(=O)C(C(O)C)NC(=O)C(C)C(O)C(C)NC(=O)C(C(OC1C(C(O)C(O)C(CO)O1)OC1C(C(OC(N)=O)C(O)C(CO)O1)O)C=1NC=NC=1)NC(=O)C1=NC(C(CC(N)=O)NCC(N)C(N)=O)=NC(N)=C1C MRNLLBXPSWMYCK-UHFFFAOYSA-N 0.000 description 1
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
- A61K31/7052—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides
- A61K31/706—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom
- A61K31/7064—Compounds having saccharide radicals and heterocyclic rings having nitrogen as a ring hetero atom, e.g. nucleosides, nucleotides containing six-membered rings with nitrogen as a ring hetero atom containing condensed or non-condensed pyrimidines
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
내용 없음.No content.
Description
제1도 및 제2도는 본 발명의 브레오마이신 NK631의 자외선 흡수곡선을 표시하는 도표.1 and 2 are diagrams showing the ultraviolet absorption curve of the breomycin NK631 of the present invention.
제3도 및 제4도는 NK631의 자외선 흡수곡선을 표시하는 도표.3 and 4 are diagrams showing ultraviolet absorption curves of NK631.
본 발명은 무독성염인 새로운 하나의 브레오마이신과 그 새로운 브레오마이신을 제조하는 방법에 관한 것이다.The present invention relates to a new bryomycin, which is a non-toxic salt, and a method for preparing the new bromycin.
더 상세히 말하면, 다음의 구조식(I)으로 표현되는 무독성염인 3-[(S)-1'-페닐에틸아미노]프로필아미노 브레오마이신과 그 새로운 브레오마이신의 제조법에 관한 것이다.More specifically, the present invention relates to a process for preparing 3-[(S) -1′-phenylethylamino] propylamino bremycin and its new breomycin, which are non-toxic salts represented by the following structural formula (I).
브레오마이신은 본 발명자중의 한사람인 우메자와 및 그 협조자[항생물에 관한 의학지 19A, 200(1966)참조]들이 발견한 가아시노다틱 항생물질로서, 액티노마이셀 스트랩트마이세스, 버디실러스로 제조 수용성 기초 그리코페프타이드이며 2가 구리의 한원자를 용이하게 키레이딩(CHELATING)할 수 있는 능력이 있다. 정상적인 배양에서 브레오마이신의 16개 성분이 생산되고 분리된다.Breomycin is a gacinodatic antibiotic discovered by one of the inventors Umezawa and its co-operators (see Antibiotics Medical Journal 19A, 200 (1966)). It is a water-soluble basic glycopeptides manufactured and has the ability to easily chelate single atoms of divalent copper. In normal culture 16 components of breomycin are produced and isolated.
[예 : 우메자의 수명·항생물지, 19A 210(1966)][Example: Life and Antibiotics of Umeza, 19A 210 (1966)]
브레오마이신 종류중에 동이 포함되지 않은 A1, A2, A5, B2, 및 디메틸 A2의 혼합물(이하 브레오마이신 복합체라 칭함)은 암치료의 임상분야에서 널리 사용되 왔으며, 주로 비늘상 세포암종, 피부암, 머리와 목의 암, 자궁암, 폐암, 악성임파선암을 치료하는데 특히 성공적이었다.Mixtures of A 1 , A 2 , A 5 , B 2 , and dimethyl A 2 (hereafter referred to as breomycin complexes), which do not contain copper among the breomycin classes, have been widely used in the clinical field of cancer treatment, mainly on scales It has been particularly successful in treating epithelial carcinoma, skin cancer, head and neck cancer, uterine cancer, lung cancer and malignant lymphadenopathy.
그러나 부작용과 관련되어 풀모노리화이브로시스와 기타 바람직하지 않은 원인 등의 사례들이 보고된 바 있다.However, there have been reports of side effects such as full monolithosis and other undesirable causes.
브레오마이신으로 인하여 나타난 부작용중에 가장 무서운 것이 풀모노리화이브로시스이다. 브레오마이신의 카아시노스타틱 작용 역할은 위에 말한 부작용이 다소간이나마 감소될 수만 있다면 임상분야에서 더 효과적으로 나타날 수 있다는 것을 용이하게 상상할 수가 있다.One of the most frightening side effects caused by breomycin is full monolithosis. It is easy to imagine that the role of breamycin in the carcinosstatic action can be more effective in the clinical field if the above-mentioned side effects can be somewhat reduced.
이러한 상황하에서 본 발명자들은 각종 브레오마이신의 합성과 주요한 시험과제로서 풀마노리화이브로시스를 포함한 그 약들의 중독성 뿐만 아니라 카시노스타틱 작용 역할에 대한 여러 가지 동물 시험에 수년간 종사하여 왔다.Under these circumstances, the present inventors have been engaged for several years in the synthesis of various breomycin and in various animal tests for the addictive as well as casinostatic action of the drugs including fulanolithosis as the main test.
그 결과로서 카복실 그룹에 브레오마이신산의 반응 유도제를 N-[(S)-1'-훼닐에틸]-1,3-디 아미노프로페인으로 반응시켜서 얻어진 새로운 브레오마이신인 3-[()-1'-훼닐에틸아미노- 프로필아미노 브레오마이신(이하 동함유 및 동비함유형을 모두 포함하여 ＂NK631＂라고 칭한다)은 상품화된 브레오마이신 복합체 및 기타 알려져 있는 브레오마이신들과 비교하여 풀마노리화이브로시스를 발생시키는 부작용을 현저하게 감소시키며 또한 카시노스타틱 작용역할을 저하시키지 않는다.As a result, 3-[()-1, a new breomycin obtained by reacting a carboxyl group with a reaction inducer of breomycin acid with N-[(S) -1'-phenylethyl] -1,3-diaminopropane '-Phenylethylamino-propylamino breomycin (hereinafter referred to as NKNK631), including both copper-containing and non-containing types,) is a full aminosylbromysis in comparison with commercialized breomycin complexes and other known breomycins. It significantly reduces the side effects that occur and also does not lower the carnostatic function.
본 발명은 위에서 말한 발견을 기초로하여 성취되었다.The present invention has been accomplished based on the above findings.
본 발명의 목적은 새로운 브레오마이신이며, 일종의 무독성염인 3-[(S)-1'-페닐에틸아미노]프로필아미노브레오마이신과 새로운 브레오마이신을 제조하는 방법을 제공하는데 있다.It is an object of the present invention to provide a process for the preparation of 3-breastin, a new non-toxic salt, 3-[(S) -1′-phenylethylamino] propylaminobreomycin and a novel brimemycin.
본 발명의 기타 목적과 장점은 다음의 설명에서 명시된다.Other objects and advantages of the invention are set forth in the following description.
구조식 (I)로서 표현되는 본 브레오마이신의 뛰어난 생화학적 작용은 시험예들과 관련하여 아래와 같이 예종된다.The outstanding biochemical action of von bremycin, represented as Structural Formula (I), is illustrated below in connection with the test examples.
＂NK631＂의 생화학적 작용할은 다음의 4개 테스트항목과 관련하여 상품화된 브레오마이신 복합체 및 3-[(R, S)-1'-페닐에칠아미노] 프로필 아미노 브레오마이신 모노설훼이트(동비함유형)(이하 ＂RS형＂이라고 칭한다)와 비교하여 실험되었다. ＂RS형＂은 미국특허 3,846,400에 발표된 발효공정으로 얻어졌다.The biochemical action of NK631 is characterized by the commercialization of breomycin complex and 3-[(R, S) -1′-phenylethylamino] propyl amino bremycin monosulfate in relation to the following four test items: It was compared with (Equivalent Type) (hereinafter referred to as "RS type"). Form RS was obtained by a fermentation process disclosed in US Pat. No. 3,846,400.
(1) 풀모나리화이 브로시스(1) Full Mona Lisa White Brosis
(2) 항종양 작용(2) antitumor action
(3) 항세균작용(3) antibacterial action
(4) 중독성(4) addictive
1. 생쥐의 폐에 대한 중독성(화이브로시스)1. Addictive to lungs of mice (fibrosis)
ICR 종족(種族) 생쥐(숫컷 생후 15주)를 구룹당 12마리씩 사용하였다.12 ICR race mice (15 weeks old) were used per group.
각 약물시험 조제약의 투여량은 5mg/kg이었다.The dose of each drug test formulation was 5 mg / kg.
투약은 1일 1회 연 10일간 복막내 주사로 투여됐다.Dosing was administered once per day by intraperitoneal injection for 10 days.
투약후 생쥐들은 5주간 양육되었다.Mice were reared for 5 weeks after dosing.
그후 관찰과 함께 그 쥐들은 살해되고 풀모나리화이브로시스의 정도를 알아내기 위하여 시체검사를 하였다.Then, with observation, the mice were killed and examined by a body to determine the degree of pulmonary phylosis.
본 발명의 새 브레오마이신과 상품화된 하나의 브레오마이신 복합제와 ＂RS형＂을 투약한 쥐에서 폴모나리화이브로시스의 발생 및 그 정도를 비교하였다.The incidence and severity of polmonalis hydrosis was compared in mice treated with the new breomycin and commercialized one breomycin combination of the present invention and "Type RS".
얻어진 결과는 표1에 표시된 바와 같다.The results obtained are shown in Table 1.
[표 1]TABLE 1
주 : *0점 : 화이브로시스 없음Note: * 0 point: No fibrosys
1점 : 알비오라 경막 알비오러스 및 화이브로시스유형의 변화에 있어서의 분피물 축적1 point: accumulation of aliquots in the change of Albiora dural Alvios and phloxis type
2점 : 수저처의 화이브로시스2 points: Cutlery
4점 : 분산된 화이브로시스4 points: distributed fibrosys
6점 : 전체의 2/3 부위에 발생된 화이브로시스6 points | pieces: The fibrosis which generate | occur | produces 2/3 of the whole part
위의 결과로부터 분명한 것과 같이 RS형 및 브레오마이신의 복합체 경우와 비교하였을 때 NK631 모노설 훼이트(등비포함형)에 기인한 화이브로시스는 발생수에 있어서 각기의 약 1/2 및 1/3로 그리고 정도에 있어서는 각기의 1/2 및 1/4로 가소되었다.As can be seen from the above results, the fibrosis attributable to NK631 monosulfite (equal inclusion type) was about 1/2 and 1/3 of the number of occurrences, as compared with the case of complexes of type RS and breomycin. And in degree, plasticized to 1/2 and 1/4 respectively.
이것은 임상분야 있어서 NK631 모노설훼이트이 유용성을 표시한다.This indicates that NK631 monosulfate is useful in the clinical field.
2. 항종양 작용2. Antitumor action
2-1 배양된 Hels S3세포에 대한 작용2-1 Action on Cultured Hels S 3 Cells
각 브레오마이신에 대한 ID50은 각 브레오마이신을 사용하면서 72시간 배양하는데 있어서의 성작 언제비율로서 계산되었다.The ID 50 for each breomycin was calculated as the ratio of when to grow in 72 hours of incubation with each breomycin.
NK 631에 대한 ID50은 브레오마이신 복합체에 대한 1.70mcg/ml에 비교하여 0.82mcg/ml임이 밝혀졌다.The ID 50 for NK 631 was found to be 0.82 mcg / ml compared to 1.70 mcg / ml for the breomycin complex.
이것은 KN 631이 브레오마이신 복합체보다 2배나 강력하게 세포성장을 억제하였음을 나타낸다.This indicates that KN 631 inhibited cell growth twice as strongly as the breomycin complex.
RS형에 대한 ID50은 0.80mcg/ml임이 밝혀졌다.The ID 50 for Form RS was found to be 0.80 mcg / ml.
이것은 그 억제효과가 NK 631 모노설훼이트의 효과와 맞먹는다는 것을 나타낸다.This indicates that the inhibitory effect is comparable to that of NK 631 monosulfate.
2-2 쥐의 애릭캔사아(Ehrlich cancer) 카시노스타틱작용(소리드튜-모-Solid tumor)2-2 Eschlich cancer casinostatic activity in rats (Sorudtu-Mo-Solid tumor)
각기 2×106세포를 ICR 종족의 쥐(6주생 숫컷)의 사타구니 부위에 피하로 이식하였다.Each 2 × 10 6 cells were implanted subcutaneously into the groin area of rats (6 week-old males) of the ICR species.
24시간후에 각 쥐는 위의 1,2항에서 설명한 바와 같은 방법으로 시험 조제약을 투여받았다.After 24 hours, each rat received the test preparation as described in
피하이식후 15일째 되던 날에 종양은 각 쥐로부터 제거하여 그 억제비율을 측정하기 위하여 치료를 받지 않은 대조구룹에서 발육된 종양의 것과 무게를 비교하엿다.On the 15th day after subcutaneous transplantation, the tumors were removed from each rat and weighed against those of the tumors developed in the untreated control group to determine their inhibition rate.
표 2에서 보는 바와 같이 NK 631 모노설훼이트의 카시노스타틱 작용은 RS형의 작용과 비슷하며 브레오마이신 복합체의 작용보다는 1.4배나 높다.As shown in Table 2, the casinostatic action of NK 631 monosulfate is similar to that of RS and 1.4 times higher than that of the breomycin complex.
[표 2]TABLE 2
2-3. 쥐내에 아싸이트스 헵파트만(아싸이트스형)에 대한 카시노스타틱 작용2-3. Casinostatic Action on Assites Heppatman (Assites) in Rats
각 1×106AH66 세포가 동류 종족쥐에게 복막내로 이식되었다.Each 1 × 10 6 AH66 cell was transplanted intraperitoneally into a cohort of rats.
24시간후에 시험 조제약은 1일 1회 연 10일간 복막내로 투약됐다.Twenty four hours later, the test formulation was administered to the peritoneum once a day for 10 days.
이식후 30일기간동안 각기 쥐에 중량과 죽은쥐와 생존쥐수가 관찰되었다.During the 30 days after transplantation, rats were weighed, dead and survived.
[표 3]TABLE 3
표 3에서 보는 바와 같이 AH66 아싸이트스 헵파트만에 대한 내성작용에서 NK 631 모노설훼이트는 RS형과 비등하며 브레오마이신 복합체에 대해서는 매투약에서 우월하였다.As shown in Table 3, NK 631 monosulfate boils over the RS form in resistance to AH66 sites heptapman and is superior to meds for breomycin complexes.
2-4 20-메틸 코란쓰린(이하 20-MC로 칭한다)에 의하여 야기된 쥐의 스크아모-스 세포 암종에 대한 억제작용 20-MC의 아세톤 용액을 원측적으로 ddy 종족의 쥐(10주생 숫컷)에 짤려진 등에 주2회 18주간 바른다.2-4 Inhibitory Effect on Scamo-S cell Carcinoma of Mice Caused by 20-Methyl Koranthrin (hereinafter referred to as 20-MC) Acetone solution of 20-MC was dominantly applied to mice of ddy species (10-year-old males). Apply twice a week for 18 weeks.
20-MC 처치시작으로부터 5주후에 62.5mcg/쥐의 시험 조제약이 주 2회 15주간 동안복막내로 투여되였다.Five weeks after the start of 20-MC treatment, a test preparation of 62.5 mcg / rat was administered intraperitoneally twice a week for 15 weeks.
시험약의 투약 완료후 첫주내에는 20-MC가 투여된 부위를 카시노제네시스의 발생을 보기 위하여 병리학적으로 검사하였다.Within the first week after the completion of dosing of the test drug, the site to which 20-MC was administered was examined pathologically to see the incidence of casinogenesis.
표 4에서 보는 바와 같이 NK 631 모노설훼이트(동 비 포함)와 RS형은 20-MC에 기인한 카시노제네시스를 억제하는데 있어서 브레오마이신 복합제보다 약간 더 효과적이다.As shown in Table 4, NK 631 monosulfate (including the same ratio) and Form RS are slightly more effective than breomycin combinations in inhibiting casinogenesis due to 20-MC.
[표 4]TABLE 4
위 결과에 나타난 바와 같이 NK 631 모노설훼이트는 상품화된 브레오마이신 복합제와 비교하여 항종양 작용이 우월함을 명백히 하였다.As shown in the above results, NK 631 monosulfate clarified superior antitumor activity compared to the commercialized breomycin combination.
3. 항균효능은 브레오마이신 A2를 표준으로 사용하므로서 사이코 박테륨 607과 바시트스 사브트레스 PCI에 대한 퀵방식으로 실험하였다. 결과는 표 5에 보이는 바와 같다.3. The antimicrobial effect was tested by the quick method for psychobacterium 607 and vacitta sabres PCI using breomycin A 2 as a standard. The results are shown in Table 5.
[표 5]TABLE 5
표 5로 명백한 것과 같이 NK 631 모노설훼이트와 RS형은 브레오마이신 복합제의 효능보다 월등한 항균효능을 보였다.As is apparent from Table 5, NK 631 monosulfate and RS showed superior antimicrobial activity than that of breomycin combination.
4. 쥐의 복막내르트(LD 50)에서의 급성 중독성4. Acute Toxicity in Rat Peritoneal Nartrum (LD 50)
표 6에서 보는 바와 같이 NK 631 모노설훼이트를 사용한 쥐에 대한 LD 50치는 브레오마이신 복합체와 RS형에 대한 것과 거이 비등하다.As shown in Table 6, LD 50 values in rats using NK 631 monosulfate are very similar to those for Bremycin complex and Form RS.
[표 6]TABLE 6
4-2 쥐와 개들의 완성 및 만성 중독4-2 Completion of Rats and Dogs and Chronic Poisoning
NK 631 모노설훼이트는 쥐와 개의 완성 및 만성 중독에 있어서 브레오마이신 복합체와 비등하다. 더욱이 주사한 곳의 탈저(necrosis)가 브레오마이신 복합제에 다량투약(1.2mg/kg 90회 주입)을 한 개의 모든 사례에서 발견되었으나 같은 량의 NK 631 모노설훼이트를 투약한 개의 사례에서는 하나도 발견되지 않았다는 사실에 대하여 특별히 말해 둔다.NK 631 monosulfate is comparable to the breomycin complex in rat and dog completion and chronic intoxication. In addition, necrosis at the site of injection was found in one case of a large dose (1.2 mg / kg 90 injections) of the breamycin combination, but not in one case of the same dose of NK 631 monosulfate. Special mention is made of the fact that it was not found.
폐에 대한 NK 631 모노설훼이트의 중독 역시 브레오마이신 복합체의 것보다 낮다. 상기 결과로 볼 때 NK 631 모노설훼이트의 중독은 폐에 대한 중독과 탈지를 제외하고는 브레오마이신 복합체의 것과 비슷하다.The intoxication of NK 631 monosulfate to the lungs is also lower than that of the breomycin complex. As a result, the poisoning of NK 631 monosulfate is similar to that of the breomycin complex except for poisoning and degreasing of the lung.
5. 결론5. Conclusion
상기한 시험결과로 볼 때 결론적으로NK 631은 다음의 특징을 갖인 새로운 화합물이다.In conclusion, NK 631 is a novel compound having the following characteristics.
(1) 폐에 대한 극히 작은 중독성(1) extremely small addictive to the lungs
(2) 상품화된 브레오마이신 복합체 보다 우월한 항균 및 항종양 작용(2) superior antimicrobial and anti-tumor activity than commercialized bromycin complexes
(3) 상품화된 브레오마이신 복합체와 비슷한 규칙적인 중독.(3) Regular addiction, similar to the commercialized breomycin complex.
(4) 주입(약물)된 곳에 대한 낮은 부분중독(4) low partial poisoning of the injected site
따라서 NK 631은 임상분야에서 유효함이 예상된다.Therefore, NK 631 is expected to be effective in the clinical field.
본 발명의 새 브레오마이신인 NK 631은 다음 구조식으로 표시되는 카복실 구름의 브레오마이신산의 반응 유도제를 N-[(S)-1'-훼닐에틸]-1.3-디 아미노프로판으로 반응시키므로서 합성한다.NK 631, a new breomycin of the present invention, is synthesized by reacting a reaction inducing agent of breamycin acid of the carboxyl cloud represented by the following structural formula with N-[(S) -1′-phenylethyl] -1.3-diaminopropane. .
더 상세히 설명하면 (1) 브레오마이신산을 N-(S)-1'-페닐에틸]-1,3-디 아미노프로판으로 반응촉진제를 사용하며 반응시키거나 (2) 브레오마이신산 3-아미노프로필에스텔 또는 N-1치환한 유도제를 N-[(S)-1'-페닐에틸]-1,3-디아미노프로판으로 반응시켜서 제조된다.More specifically, (1) breomycin acid may be reacted with N- (S) -1'-phenylethyl] -1,3-diaminopropane with a reaction accelerator or (2) breomycinic acid 3-aminopropyl It is prepared by reacting an ester or an N-1 substituted derivative with N-[(S) -1'-phenylethyl] -1,3-diaminopropane.
이들 공정의 상세한 내용은 아래와 같다.Details of these processes are as follows.
공법(1)에서 사용된 브레오마이신산은 미국 특허 3,843,448호 및 3,846,400호에 발표된 방법에 따른 브레오마이신 B2의 효소분해로서 얻어진 기히 알려진 화합물이다.Bremycin acid used in process (1) is a well known compound obtained by enzymatic digestion of breomycin B 2 according to the methods disclosed in US Pat. Nos. 3,843,448 and 3,846,400.
또 하나의 사용물질 즉 N-[(S)-1'-페닐에틸]-1,3-디 아미노프로판은 다음 방식으로 본 발명자등이 처음 합성한 새로운 화합물이다.Another useful substance, N-[(S) -1'-phenylethyl] -1,3-diaminopropane, is a new compound first synthesized by the present inventors in the following manner.
약 0℃에서 냉각된 페닐에틸 아민을 대략 같은 량의 아크리토니트릴과 혼합한다.Phenylethyl amine cooled at about 0 ° C. is mixed with an approximately equal amount of acritonitrile.
혼합물은 반응을 완성하도록 8℃∼100℃에서 10-24시간 동안 유지한다. 초과 아크릴로니트릴은 잠압증류로 제거하고, 잔사는 3-[(3)-1'-페닐에틸아미노] 프로피오리트릴을 얻도록 더 증류한다. 이렇게하여 얻은 화합물은 관습적인 방식으로 예를 들면 목적한 N-[(S)-1'-페닐에틸]-1,3-디 아미노프로판(이하 아미노 화합물이라 칭한다)를 산출하도록 레니·니켈을 사용하여 분해한다.The mixture is held at 8 ° C.-100 ° C. for 10-24 hours to complete the reaction. Excess acrylonitrile is removed by latent distillation, and the residue is further distilled to obtain 3-[(3) -1'-phenylethylamino] propioritryl. The compound thus obtained is used in a customary manner, for example, by using nickel to yield the desired N-[(S) -1'-phenylethyl] -1,3-diaminopropane (hereinafter referred to as amino compound). Disassemble it.
이 화합물에 물리화학적 특징은 표 7에 기재된 바와 같다.The physicochemical characteristics of this compound are shown in Table 7.
[표 7]TABLE 7
유용한 반응촉진제로 예를 들면, 6-크로로-1-P-크로로벤젠설휘닐 옥시벤조트리아졸(CCBT), N-에틸-5-페닐 이소자조리움-3'-설화네이트(NEPIS), N-터트-부틸-5-메틸이소자조리움 과염소산염류 N-에톡시-카보닐-2-에톡시-1,2-디 하이드로퀴노린, 디-P-니트로페닐설화이트, 트리-P-니트로페닐 인산염, P-니트로페닐 트리크로로아세테이트, N-하이드로키시 서시니마이드, 디싸이크로헥실카보디마이드[DCC], 1-에틸-3-(3-디메틸아미노푸로필)-카보디마이드, 1-싸이크로헥실-3-(2-몰호리노에틸)카보디마이드, 디페닐카보디마이드, 디-P-토루일카보디마이드, 디 이소푸로필카보디마이드, P-니트로페놀, 펜타크로로페놀, 벤질알콜 등을 들 수 있다.Useful reaction promoters include, for example, 6-chloro-1-P-chlorobenzenesulfonyl oxybenzotriazole (CCBT), N-ethyl-5-phenyl isozorium-3'-sulfonate (NEPIS), N-tert-butyl-5-methylisozazolium perchlorate N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline, di-P-nitrophenylsulwhite, tri-P-nitro Phenyl phosphate, P-nitrophenyl trichloroacetate, N-hydrokisyl sercinimide, dicyclohexylcarbodiamide [DCC], 1-ethyl-3- (3-dimethylaminofurophyll) -carbodimide, 1-cyclohexyl-3- (2-molhorinoethyl) carbodimide, diphenylcarbodimide, di-P-tolylcarbodimide, diisopurophyllcarbodimide, P-nitrophenol, pentacrorophenol And benzyl alcohol.
공정의 상세한 설명(1) :Detailed description of the process (1):
브레오마이신산(동포함형)은 물, 디메틸호름마이드, 디메틸설화옥사이드, 또는 그의 혼합물에서 용해한다.Breomycinic acid (inclusive) dissolves in water, dimethylhommide, dimethyl sulfide oxide, or mixtures thereof.
그 용액에 상기한 반응 촉진제중의 하나를 첨가하고 0-30℃에서 교반한다.One of the reaction promoters described above is added to the solution and stirred at 0-30 ° C.
얻어진 용액을 염산 또는 수산화 나트륨과 같은 무기산이나 염기를 첨가하거나 또는 트리크로로아세틱에시드 혹은 N-메틸몰호린 등과 같은 유기산이나 염기를 첨가하여 반응촉진에 적당한 pH3-10으로 조정한다. 이러한 처리를 함으로서 브레오마이신산의 카복실구룹 반응이 시작된다.The obtained solution is adjusted to pH 3-10 suitable for promoting the reaction by adding an inorganic acid or base such as hydrochloric acid or sodium hydroxide, or an organic acid or base such as trichloroacetic acid or N-methylmolhorin. This treatment initiates the carboxyl group reaction of breomycin acid.
처리후 즉시 또는 30분 이내에 반응용액을 약 pH7.0으로 조정하는 동안 또는 그후에 아미노 화합물(위에 말한 바 있음)을 첨가 혼합한다. 이 혼합물은 반응이 진행되게 하기 위하여 ℃-30℃에서 1-24시간동안 유지하여 NK 631를 산출한다.Immediately or within 30 minutes after treatment the amino compound (as mentioned above) is added and mixed during or after adjusting the reaction solution to about pH7.0. The mixture is held at 1-230 C for 1-24 hours to allow the reaction to proceed to yield NK 631.
반응촉진제 또는 아미노화합물의 적당한 비율은 브레오마이신산 당량에 대하여 1 내지 10당량의 비율이다.A suitable ratio of the reaction accelerator or amino compound is 1 to 10 equivalents relative to the equivalent of bromycin acid.
반응 혼합물에서 NK 631을 분리하기 위하여 먼저 브레오마이신 화합물을 그 반응혼합체의 1분량에 대하여 아세톤 5-10의 분량을 첨가하여 철저하게 침전시킨다.In order to separate NK 631 from the reaction mixture, the bromycin compound is first thoroughly precipitated by adding an amount of acetone 5-10 to 1 part of the reaction mixture.
그 침전물은 여과수집하고 아세톤으로 세척하며 가능한한 최소량의 증류수에 용해시킨다.The precipitate is collected by filtration, washed with acetone and dissolved in the smallest possible amount of distilled water.
그 수성용액은 즉시 염산 또는 비슷한 종류의 약품으로 pH60으로 즉시 조정하여 수성염화암모니아 용액으로 포장된 CM-Sephdex C-25(NH4 +형으로서 화-마샤화인케미칼사의 제품임)가 들어 있는 크로모도크라휘카람위로 주입된다.The aqueous solution is immediately chromodome containing CM-Sephdex C-25 (NH 4 + type, manufactured by Hwa-Masha Fine Chemicals) packaged in aqueous ammonia chloride solution, immediately adjusted to
미반응의 브레오마이신산은 그 카람을 흡착되지 않이하고 통과한다. 수지에 흡착된 NK 631은 0.05M에서 1.0M로 단계적 또는 계속적으로 증가하는 제조농도의 수성염화암모니아 용액이 용출된다.Unreacted bromycin acid passes through the caram without adsorption. The NK 631 adsorbed on the resin is eluted with an aqueous ammonia chloride solution having a manufacturing concentration increasing stepwise or continuously from 0.05M to 1.0M.
NK631은 292mμ에서 자외선 흡착을 나타내는 염화암모니아 농도 약 0.35M∼0.45M에서 미세한 청색유출물속에 포하되 있다. 이 유출물은 분리수집하여 Amberirte
이 유출물에서 순수하고 청색의 비결정 분말 NK 631(동 포함형)을 얻는다.From this effluent, pure and blue amorphous powder NK 631 (copper containing type) is obtained.
NK 631중의 염화수소 및 유화물은 각각 염산 및 황산을 포함하는 수성 메타놀로 세척하여 얻는다.Hydrogen chloride and emulsions in NK 631 are obtained by washing with aqueous methanolol containing hydrochloric acid and sulfuric acid, respectively.
이 공정에서 미 반응의 브레오마이신산은 용이하게 회수할 수 있다.In this process, unreacted bromycin acid can be easily recovered.
CM-Sephadex
예를 들면 수성염화나트륨 또는 수성 황산 암모니아가 세척제로서 사용될 수 있다.For example, aqueous sodium chloride or aqueous ammonia sulfate may be used as the cleaning agent.
위에서 얻은 동을 포함하는 NK 631로부터의 동제거는 동이 침전되는 황산동으로 변형 전환시키기 위하여 유화수소를 반응시키는 방법.Copper removal from NK 631 comprising copper obtained above is a method of reacting hydrogen emulsification to transform the copper into precipitated copper sulfate.
환원제로 동을 0원자가 구리로 환원시키는 방법(미특허 3646197호)디치죤[우메자와외 항생물질지 19A, 210(1960)]과 같은 킬레이팅 작용제를 포함하는 유기용제로 구리를 추출하는 방법 또는 비 이온교환 마크로레티크라 레-진을 사용하는 방법(미특허 3929993호)중 어느 것이 적용하여 달성할 수 있다.A method of extracting copper with an organic solvent containing a chelating agent such as dizijon (Umezawa et al. 19A, 210 (1960)), or a method of reducing copper to zero-valent copper with a reducing agent or non-pattern copper (US Pat. No. 3646197). Any of the methods using ion exchange macroreticra resin (US Pat. No. 3,929,993) may be applied and achieved.
구리제거공정의 한 예를 아래와 같이 상술한다.An example of a copper removal process is explained in full detail below.
NK 631(구리포함형)을 증류수에 용해시키고 그 용해액을 NK 631이 흡착되게 하기 위하여 증류수에 포장된 Amberli-te
(Diaion
그 다음 그 카람을 에틸렌 디 아민 테트라 아세틱에서도 피소듐(이하 EDTA. Na2로 칭한다)의 5% 수용액으로 세척한다.The caram is then washed with 5% aqueous solution of physodium (hereinafter referred to as EDTA. Na 2 ) also in ethylenediamine tetraacetic.
그러며는 동 이온은 EDTA. Na2에 의하여 분리제거되고 수지상태는 동을 포함하지 않는 NK 631만 남는다.The copper ion is EDTA. It is separated off by Na 2 and the resin remains only NK 631 which does not contain copper.
그 수지는 염화나트륨, 황산나트륨, 초산나트륨과 같은 염의 수용액으로 세척하여 EDTA-Na2를 제거하고나서 증류수로 세척한다. 끝으로 예를 든다면, 페타놀과 0.0025N 염산(1 : 1V/V)의 혼합물과 같은 산성 메타놀-물의 혼합물을 카람에 통과시켜 290m에서 자외선 흡수를 보이는 물질을 세척해낸다.The resin is washed with an aqueous solution of a salt such as sodium chloride, sodium sulfate, sodium acetate to remove EDTA-Na 2 and then washed with distilled water. Finally, for example, a mixture of acidic metaol-water, such as a mixture of petanol and 0.0025N hydrochloric acid (1: 1V / V), is passed through the caram to wash away materials that exhibit ultraviolet absorption at 290 m.
이 물질을 수집, 농축하여 PH 6.0으로 조정하고 엷은 황백색의 비결정분말인 NK 631 디하이드로크로라이드(구리비포함형)을 산출하도록 냉동 건조한다.This material is collected, concentrated to pH 6.0, and freeze-dried to yield NK 631 dihydrochloride (copper free), a pale yellow-white amorphous powder.
산성 메타놀-물의 혼합물을 조제하는데 사용될 산으로는, 의약적으로 허용되는 한 아무 산이나 사용할 수 있다.As the acid to be used for preparing the acidic methanol-water mixture, any acid can be used as long as it is pharmaceutically acceptable.
예를 들면, 황산이나 초산을 사용하며는 NK 631 1화황산염 또는 2화초산염(구리비포함형)이 얻어진다.For example, using sulfuric acid or acetic acid, NK 631 monosulfate or diacetate (copper-free) is obtained.
본 발명의 NK 631은 보통의 방법으로 기타 무독성염인 황산 및 초산염으로 변형할 수 있으며, 그것은 세척해 사용된 산을 임의 변경함으로서 이룰 수 있다.The NK 631 of the present invention can be modified in the usual way with other non-toxic salts, sulfuric acid and acetate, which can be achieved by washing and arbitrarily changing the acid used.
공정(1)에서는, 그 아미노화합물이 포리헵타이드합성에 사용된 반응촉진제의 조재로 브레오마이신산의 활성화된 카복실구룹과 반응하기 때문에 축합반응은 극히 순한상태에서 원활하게 진행된다. 결과적으로 아미노화합물중의 1차 아미노구룹만이 활성화된 카복실구룹과 반응하며 2차 아미노구룹은 그 동안에 반응에 참여하지 아니한다. 이렇게하여 NK 631이 우선적으로 산출된다. 이것이 본 공정법(1)의 뛰어난 점이다.In step (1), the condensation reaction proceeds smoothly in an extremely mild state because the amino compound reacts with the activated carboxyl group of bromycin acid with the aid of the reaction promoter used in the synthesis of polyheptide. As a result, only the primary amino group in the amino compound reacts with the activated carboxyl group and the secondary amino group does not participate in the reaction during that time. In this way, NK 631 is preferentially calculated. This is the outstanding point of this process method (1).
공정의 상세한 설명(2) :Detailed description of the process (2):
공법(2)에 사용된 브레오마이신산의 3아미노푸로필에스텔은 3-(메칠멜카프토)푸로필 아미노브레오마이신을 형성하도록 브레오마이신 A2를 가열분해시켜, 거기서 나오는 브레오마이신을 산용액에서 할로겐니트릴, 할로겐초산, 할로겐초산염에스텔 또는 할로겐아세트아미드로 반응시켜서 디하이드로크로라이드의 형태로 용이하게 얻을 수 있다.The 3 aminofurophylester of breomycin acid used in the process (2) thermally decomposes breomycin A 2 to form 3- (methylmelcapto) furophyl aminobreomycin, and the resulting broomycin in acid solution. It can be easily obtained in the form of dihydrochloride by reaction with halogen nitrile, halogen acetate, halogen acetate ester or halogen acetamide.
자체의 아미노구룹을 보호하여 갖고 있는 브레오마이신산중의 3-아미노푸로필 에스텔의 N-1가치환한 유도제는 다음과 같이하여 다량 산출한다.The N-1 substituted derivative of 3-aminofurophyl ester in bromycin acid which protects its own amino group is computed in large quantities as follows.
먼저 브레오마이신산중의 3-아미노푸로필 에스텔의 디하이드로크로라이드를 물이나 또는 메타놀 혹은 그 혼합물 등의 유기용제에 용해시키고나서, 그 용액에 분말형태아 메타놀과 같은 유기용제에 용해된 형태인 기존 아미노구룹 보존반응제중의 하나를 같은 분량 또는 약간 초과하는 량으로 강력하게 교반과 함께 서서히 첨가하고, 반응혼합물의 pH를 예를 들면 트리메틸아민, 트리에틸아민, 피리딘, 1,3-디아자비싸이크로-[5,4,0]-7-언데씬, 1,5-디아자비싸이크로-[3,4,0]-노년, 1,4-디아자비싸이크로-[2,2,2]-옥탄, 또는 N-메틸몰포린 등과 같은 유기염기를 첨가하여 5.0 내지 7.5로 계속유지하면서 실내온로 혹은 빙냉 온도에서 반응이 진행되게 한다.First, the dihydrochromide of 3-aminofurophil ester in bromyceic acid is dissolved in an organic solvent such as water or methanol or a mixture thereof, and then dissolved in an organic solvent such as powdered methanol in the solution. One of the existing aminogroup preservatives is added slowly with vigorous stirring in the same or slightly above amount and the pH of the reaction mixture is for example trimethylamine, triethylamine, pyridine, 1,3-diazabi Micro- [5,4,0] -7-unscene, 1,5-diazabicyclo- [3,4,0] -old age, 1,4-diazabicyclo- [2,2,2 ] -Octane, or an organic base such as N-methylmorpholine is added to keep the reaction proceeding at room temperature or ice-cooling temperature while continuing to 5.0 to 7.5.
그러한 N-일치환된 유도제중의 대표적인 것이 다음과 같다.Representative of such N-substituted inducers are as follows.
브레오판이신산중의 3-아세틸아미노프로필에스텔의 모노하이드로크로라이드, 브레오마이신산의 3-써씨닐아미노프로필에스텔, 브레오마이신산중의 3-벤조일 아미노프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-벤지톡시카보닐 아미노프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-P-토루엔설호닐아미노프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-(2,4-디니트로페닐) 아미노프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-(3,5-디메틸-3-옥소싸이크로핵젠-1-일) 아미노프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-N-터트-부톡시카보닐프로필 에스텔의 모노하이드로크로라이드, 브레오마이신산중의 3-N-사리실리딘아미노푸로필 에스텔의 모노하이드로크로라이드, (미국특허 3,886,133) 공정12)에서, NK 631은 부레오마이신산중의 3-아미노프로필 에스텔이나 N-1가치환된 3-아미노프로필 에스텔과 아미노화합물간의 반응으로서 형성된다.In monohydrochloride of 3-acetylaminopropyl ester in brenophanic acid, 3-hydrocinylaminopropyl ester of bromycinic acid, monohydrochloride of 3-benzoyl aminopropyl ester in bromycinic acid, and bromycinic acid Monohydrochloride of 3-benzithoxycarbonyl aminopropyl ester of, monohydrochloride of 3-P-toluenesulfonylaminopropyl ester in bromycin acid, 3- (2,4- in bromycin acid Monohydrochloride of dinitrophenyl) aminopropyl ester, monohydrochloride of 3- (3,5-dimethyl-3-oxocyclonugen-1-yl) aminopropyl ester of bromycinic acid, bromycinic acid Monohydrochloride of 3-N-tert-butoxycarbonylpropyl ester in water and monohydrochloride of 3-N-sarisilidineaminofurophyll ester in bromycinic acid In Id, (U.S. Patent 3,886,133) Step 12), NK 631 is formed as a reaction between 3-aminopropyl ester, or N-1 value unsubstituted 3-amino-propyl ester and amino compound in the Oh, my bladder acid.
처음에 사용하는 물질로서의 브레오마이신산의 N-1 치환된 3-아미노프로필 에스텔의 경우, 브레오마이신산의 3-아미노프로필 에스텔로부터 농축하여 얻은 순수한 물질 혹은 반응혼합물과 아미노구를 보존반응제를 사용할 수 있다.In the case of N-1 substituted 3-aminopropyl ester of breomycin acid as the first substance to be used, a pure substance or reaction mixture and amino sphere obtained by concentrating from 3-aminopropyl ester of bleomycin acid may be used as a preservative agent. Can be.
브레오마이신산의 3-아미노프로필 에스텔이나 N-1 치환된 3-아미노프로필 에스텔과 아미노화합물의 반응에 쓰이는 바람직한 용제는 물과 메타놀, 디메틸호름마이드, 디메틸 설프옥사이드 등의 유기용제이다.Preferred solvents for the reaction of the 3-aminopropyl ester of bromymic acid or the N-1 substituted 3-aminopropyl ester with an amino compound are organic solvents such as water, methanol, dimethyl-hormamide and dimethyl sulfoxide.
반응혼합물은 중성 또 아르카리상태에서 0℃-80℃에서 1-72시간 동안 유지하여 아미노리시스반응이 진행되게하여 NK 631을 형성케 한다.The reaction mixture is maintained for 1-72 hours at 0 ° C.-80 ° C. in the neutral or alkaline state to allow the aminosis reaction to proceed to form NK 631.
pH가 보다 높은 상태에서는 비교적 저온도에서 더 연장된 반응시간이 바람직하며, pH가 더 낮은 상태에서는 고온에서 짧은 반응시간이 바람직하다.Longer reaction times at relatively low temperatures are preferred at higher pH, and shorter reaction times at higher temperatures are preferred at lower pH.
반응혼합물중의 아미노화합물의 적정비율은 브레오마이신산의 3-아미노프로필에스텔 또는 까은산의 N-일치환된 유도제의 1당량에 대하여 1-10당량에 해당하는 비율이다.The proper ratio of amino compound in the reaction mixture is 1-10 equivalents to 1 equivalent of N-monosubstituted inducer of 3-aminopropyl ester of bromycinic acid or of the black acid.
반응혼합물롭터 NK 631를 분리하기 위하여 먼저 브레오마이신성분을 반응혼합물의 2 내지 5뱅 해당하는 불량 아세톤을 첨가하여 철저하게 침전시킨다.In order to separate the reaction mixture filter NK 631, first, the bromycin component is thoroughly precipitated by adding the poor acetone corresponding to 2 to 5 banks of the reaction mixture.
침전물을 여과수집하여 아세톤으로 철저하게 세척하고나서 가능한한 최소불량의 증류수에 용해한다.The precipitate is collected by filtration, washed thoroughly with acetone and dissolved in the least amount of distilled water as possible.
그 수성용액을 염산이나 그와 유사한 물질을 첨가하여즉시 pH 6.0으로 조정하고 0.05M 수성염화 암모니아 용액에 채워 담은 CM-Sephaex
그 카람을 통과함에 따라 수성염화암모니아 용액은 농도가 0.05M에서 1.0M까지 단계적 또 계속적으로 증가한다.As the caram passes, the aqueous ammonia chloride solution increases step by step continuously from 0.05M to 1.0M.
브레오마이신산의 미반응된 3-아미노프로필에스텔이나 N-일치환된 유도제와 NK 631은 염화암모니아 농도를 각각 0.15-0.20M와 0.35-0.45M에서 유출유체내에 청색반드(자외선흡수 292mμ에서 함)를 형성하며 정제된다.Unreacted 3-aminopropylester or N-monosubstituted inducer of breomycin acid and NK 631 have blue bands in the effluent fluid at 0.15-0.20M and 0.35-0.45M, respectively (with UV absorption at 292mμ). It is formed and purified.
이들 미세물을 별도로 수집하여 Amberlite
염화수소와 유화물은 각각 염산과 황산으로 산화된 물-메타놀 혼합물을 용리액으로 사용하여 얻는다.Hydrogen chloride and emulsions are obtained by using a water-methanol mixture oxidized with hydrochloric acid and sulfuric acid, respectively, as eluent.
브레오마이신산의 반응되지 않은 3-아미노프로필 에스텔이나 N-일치환의 유도제는 용이하게 회수될 수 있다.The unreacted 3-aminopropyl ester of N or unsubstituted bromine may be readily recovered.
CM-Sephadex
공법 (2)로 하여 얻은 NK 631은 공법(1)과 관련하여 전술한 기존 구리제거법을 적용하여 구리없는 형으로 변질시킬 수 있다. 필요하면, 그것을 예로 염화수소 유화물초산염등과 같이 다른 무독염으로 변질시키는 것도 가능하다.The NK 631 obtained by the process (2) can be changed into a copper-free form by applying the conventional copper removal method mentioned above with respect to the process (1). If necessary, it is also possible to alter it with other non-toxic salts such as, for example, hydrogen chloride sulphate acetate.
NK 631의 주요 물리화학적 특징들은 표 8 및 9에서 보는 바와 같다.The main physicochemical characteristics of NK 631 are shown in Tables 8 and 9.
NK 631의 구조식(1)은 다음 방법으로 확인하였다.Structural formula (1) of NK 631 was confirmed by the following method.
NK 631 모노설화이트(구리비포함형)을 증류수에 용해하여 국내기준으로 디옥산을 사용하는 푸토트늄의 축적흡수 방법으로 13C-NMR을 측정하였다.NK 631 monosulfate (copper-free type) was dissolved in distilled water and 13C-NMR was measured by accumulation absorption method of futonium using dioxane as a national standard.
측쇄 및 3-[(S)-1'-훼닐에틸아미노] 프로필아미노 반분과 함께 아미노화합물에 포함된 총제 11개의 13C 원자로 인한 신호감도가 19.4, 26.3, 37.0, 43.5, 58.9, 128.3(2개 신호), 130.0(2개신호), 136.31,ppm에서 인정되었다.Signal sensitivity due to a total of 11 13C atoms contained in the amino compound together with the side chain and 3-[(S) -1'-phenylethylamino] propylamino half, 19.4, 26.3, 37.0, 43.5, 58.9, 128.3 (2 signals ), 130.0 (2 signals), 136.3 1 , ppm.
기타 탄소원자들로 인한 신호는 브레오마이신족에 공통된 신호에 모두 부합되었다.The signals from other carbon atoms matched all of the signals common to the Breomycins.
[항생물질지, 30, 388(1977) 나가나와의][Antibiotics, 30, 388 (1977) in Naganawa]
[표 8]TABLE 8
[표 9]TABLE 9
도면중 도표 1과 2는 본 발명이 브레오마이신 NK 631의 자외선 흡수곡선을 나타내고 도표 3과 4는 NK 631의 자외선 흡수곡선을 나타낸다.In the drawings, Tables 1 and 2 show the ultraviolet absorption curves of breomycin NK 631, and Tables 3 and 4 show the ultraviolet absorption curves of NK 631.
즉, 도표 1은 3-[(S)-1'-페닐에틸아미노] 프로필 아미노 브레오마이신 디 하이드로 크로라이드(구리포함형)의 자외선 흡수곡선을 나타내며 도표 2는 3-[(S)-1'-훼닐에틸아미노] 프로필아미노 브레오마이신 모노설훼이트(구리비포함형)의 자외선 흡수 곡선을 나타낸다.That is, Table 1 shows the ultraviolet absorption curve of 3-[(S) -1'-phenylethylamino] propyl amino bremycin dihydro chromide (copper containing) and Table 2 shows 3-[(S) -1 Ultraviolet absorption curves of '-phenylethylamino] propylamino breomycin monosulfate (copper not included).
도표 3은 취화가리 정의 형태로 측정한 3-[(S)-1'-훼닐에틸아미노] 프로필아미노 브레오마이신 디하이드로 크로라이드(구리포함형)의 적외선 흡수곡선을 나타내고 도표 4는 취화가리 정형태로 측정한 3-[(S)-1'-훼닐에틸아미노] 프로필 아미노 브레오마이신(구리비포함형)의 적외선 흡수곡선을 보인다.Table 3 shows the infrared absorption curves of 3-[(S) -1'-phenylethylamino] propylamino bleomycin dihydro chromide (copper included) measured in the form of embrittlement, and Table 4 is the embrittlement definition. Infrared absorption curve of 3-[(S) -1′-phenylethylamino] propyl aminobreomycin (copper not included) measured by.
본 발명은 실시예에 의하여 상세하게 선명하면 다음과 같다.When the present invention is clear in detail by the embodiment as follows.
그러나 본 발명은 그 예들에 한정된 것은 아니다.However, the present invention is not limited to the examples.
[실시예 1]Example 1
NK 631의 디하이드로크로라이드(구리포함형) 및 모노설훼이트(구리비포함형)의 합성.Synthesis of Dihydrochloride (copper containing) and Monosulfate (copper free) of NK 631.
A공정 :A process:
디메틸호름 아마이드 400ml에 브레오마이신산(구리포함형) 15.0g을 용해한다. 그 용액에 냉각하여 0℃로 유지되고 있는 동안 N-메틸 모포린 1.1ml과 CCBT 10.3g을 첨가한다. 그 혼합물을 0.℃에서 5분간 교반한 후, 아미노 화합물 5.3g을 첨가 혼합한다.Dissolve 15.0 g of breomycinic acid (copper-containing) in 400 ml of dimethylhormide. To the solution is added 1.1 ml of N-methyl morpholine and 10.3 g of CCBT while cooling and holding at 0 ° C. After stirring the mixture for 5 minutes at 0.degree. C., 5.3 g of the amino compound is added and mixed.
그리고 1시간동안 더 교반한다.And stir for an additional hour.
25% 수성 초산 용액 200ml를 가한 반응이 끝난 후에 그 반응 혼합물은 반의 생성물을 침전시키도록 차거운 아세톤 5ℓ와 혼합한다. 침전물은 여과수집하여 아세톤으로 세척하고 증류수 500ml체 용해한다.After completion of the reaction with 200 ml of 25% aqueous acetic acid solution, the reaction mixture is mixed with 5 liters of cold acetone to precipitate half the product. The precipitate is collected by filtration, washed with acetone and dissolved in 500 ml of distilled water.
이렇게 하여 얻은 용액은 즉시 pH6.0으로 조정하고 브레오 마이신을 흡수하도록 0.05M 수성염화암모니아 용액에 담겨진 CM-Sephadex
수성 염화암모니아 용액을 사용하여 여화암모니아 농도가 0.05에서 1.0M까지 계속적으로 증가되는 카람에 용리액 20ℓ를 통과시키므로서 정제가 이루어진다.Purification is carried out using an aqueous ammonia chloride solution by passing 20 liters of eluent through a caram which continuously increases the ammonia concentration from 0.05 to 1.0 M.
미반응의 브레오마이신산은 암모니아 크로라이드 농도 약 0.05M에서 유출물이 존재하며 NK 631은 약 0.45M의 농도에 염화암모늄에 존재한다.Unreacted bromycin acid is present in the effluent at ammonia chromide concentration of about 0.05M and NK 631 is present in ammonium chloride at the concentration of about 0.45M.
292mμ에서 자외선 흡수를 보이는 그 두 물질은 별도로 각각 수집한다.The two materials, which show ultraviolet absorption at 292 mμ, are collected separately.
NK631를 포함하는 물질은 Amberlite
용리유분에서 메타놀을 증발시킨후 농축물은 Dowex
비슷한 처리로 미반응의 브레오마이신산(구리 포함형) 280mg을 회수하였다.Similar treatment recovered 280 mg of unreacted bromycinic acid (copper included).
NK 631 하이드로크로라이드(구리포함형)의 자외선 흡수 및 항균효능은 아래에 보는 바와 같다.Ultraviolet absorption and antimicrobial activity of NK 631 hydrochloride (copper included) is shown below.
자외선 흡수 최대치UV absorption maximum
[주 : 항균효능은 시험 생물로서 마이코박테륨 스매그매티스(Mycobacteriun smegmatis)[Note: The antimicrobial effect is Mycobacteriun smegmatis as a test organism.
ATCC 607를 사용하고 브레오마이신 A2(동 비포함형)의 효능을 1,000u/mg로 가정하여 시험하였다. 이하 동일하게 적용된다.It was tested assuming ATCC 607 and the efficacy of breomycin A 2 (not included) was 1,000 u / mg. The same applies to the following.
B공정 :B process:
증류수 200ml에 NK 631 하이드로크로라이드(구리포함형) 10.0g을 용해 시켰다. 그 용액을 증류수에 담은 Amberlite
그리고나서 그 카람 EDTA,Na25%를 포함하는 수성용액 2ℓ와 5% 황산나트륨요액 2.5ℓ 그리고 증류수 630mℓ로 차례차례 세척하였다.It was then washed sequentially with 2 liters of aqueous solution containing Karam EDTA, Na 2 5%, 2.5 liters of 5% sodium sulfate solution and 630 ml of distilled water.
그다음에 그카람을 메타놀-물 혼합물 (1:1V/V)에 탄 0.0025N 황산으로 침적시켰다.The caram was then deposited with 0.0025 N sulfuric acid burnt in a methanol-water mixture (1: 1 V / V).
290mμ에서 자외선 흡수를 보인 한 물질을 포함하는 미세물질 총계 900ml를 수집하였다. 증류로 메타놀을 제거한 후 잔액을 Dowex
이 생성물은 아래와 같은 자외선흡수 최대치와 항균효능을 보였다.This product showed the following maximum UV absorption and antimicrobial effect.
자외선 흡수최대치UV absorption maximum
NK 631 디하이드로크로라이드(동포함형)의 합성 증류수 40ml에 브레오마이신산(동포함형) 1.5g을 용해하였다. 26℃에서 교반중인 용액에 반응물질의 pH를 0.1N의 수산화나트륨용액을 가하여 0.8-5.5로 유지하면서 NEPIS 800mg을 첨가하였다. 30분후에 그 혼합물에 증류수 200ml의 아미노화합물 1.8g을 용해 시키고 염산으로 pH 7.0에 조절하여 조제한 용액을 첨가하였다.Synthesis of NK 631 Dihydrochloride (included) 1.5 g of breomycinic acid (included) was dissolved in 40 ml of distilled water.
그 혼합룰은 26℃에서 20시간동안 방치되였다.The mixing rule was left at 26 ° C. for 20 hours.
그 반응 혼합물에 브레오마이신 성분을 침전시키도록 차거운 아세톤 60ml를 교반하면서 참가하였다.The reaction mixture was stirred with stirring 60 ml of cold acetone to precipitate the breomycin component.
그 침전물을 여과 수집하여 아세톤으로 세척하고 증류수 30ml에 용해하였으며 즉시 0.1N 염산으로 pH 6.0으로 조절하였다. 그리하여 얻은 수용액은 실시예 1의 A공법에서 설명한 공법에 따라 점화하여 항균효능 8,073u/mg을 갖은 청색비결정 분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 932mg(53% 수확)을 얻었다.The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water, and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid. The aqueous solution thus obtained was ignited in accordance with the method described in method A of Example 1 to obtain 932 mg (53% harvest) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,073 u / mg.
[실시예 3]Example 3
NK 631 디하이드로크로라이드(구리포함형)의 합성 디메틸 설프옥사이드 40ml의 브레오마이신산(구리포함형) 1.5g을 용해하였다. 30℃에서 교반중인 용액에 N-에톡시카보닐-2-에톡시-1,2-디하이드로퀴노린 740mg와 N-메틸모포린 0.1ml를 첨가하였다.Synthesis of NK 631 Dihydrochloride (Copper Type) 1.5 g of bromycinic acid (Copper Type) of 40 ml of dimethyl sulfoxide was dissolved. To the stirring solution at 30 ° C., 740 mg of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline and 0.1 ml of N-methylmorpholine were added.
30분간 교반된 그 혼합물에 아미노 화합물 530mg을 교반하면서 첨가하였다. 그리고 그 혼합물은 30℃에서 3시간동안 방치하였다. 그 반응 혼합물에 차거운 아세톤 600ml를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다.To the mixture was stirred for 30 minutes, 530 mg of amino compound was added with stirring. And the mixture was left at 30 degreeC for 3 hours. 600 ml of cold acetone was added to the reaction mixture with stirring to precipitate the breomycin component.
그 침전물을 여과수집하여 아세톤으로 세척하고 나서 증류수 30ml에 용해하여 즉시 0.1N 염산으로 pH6.0에 조절하였다. 그리하여 얻은 수용액을 실시예 1의 A 공정에 있는 방법과 같은 식으로 처리하여 항균효능 8,028u/mg를 나타내는 청색 비결정 분말형태의 NK 631 디하이드로크로라이드(구리포함형) 809mg(46% 수확)을 얻었다.The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water, and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid. The aqueous solution thus obtained was treated in the same manner as in the process A of Example 1 to obtain 809 mg (46% yield) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,028 u / mg. Got it.
[실시예 4]Example 4
NK 631 디하이드로크로라이드(구리포함형)의 항상증류수 40ml에 브레오마이신산(구리포함형) 1.5g을 용해하였다.1.5 g of breomycinic acid (copper-containing) was dissolved in 40 ml of distilled water of NK 631 dihydrochloride (copper-containing).
27℃에 1-에틸-3-(3-디메틸 아미노프로필)-카버디이미드 하이드로크로라이드 1.2g을 첨가한 후 그 혼합물을 0.1N 염산으로 pH4.5에 조절하였다.1.2 g of 1-ethyl-3- (3-dimethyl aminopropyl) -carverimide hydrochloride was added at 27 DEG C, and the mixture was adjusted to pH 4.5 with 0.1 N hydrochloric acid.
아미노 화합물 530mg을 첨가한 후 그 혼합물을 1N 염산으로 pH 5.0에 다시 조절하고 27℃에서 20시간동안 방치하였다. 그 반응 혼합물에 차거운 아세톤 400ml를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다.After addition of 530 mg of amino compound, the mixture was adjusted again to pH 5.0 with 1N hydrochloric acid and left at 27 ° C. for 20 hours. 400 ml of cold acetone was added to the reaction mixture with stirring to precipitate the breomycin component.
그 침전물을 여과수집하여 아세톤으로 세척후 증류수 30ml에 용해하고 0.1N 수산화나트륨으로 pH 6.0에 즉시 조절하였다. 이렇게하여 얻은 수용액을 실시예 1의 A 공정에 상술한 공법에 따라 정제하여 항균효능 7,985u/mg를 갖인 청색 비결정 분말형태의 NK 631 디하이드로크로라이드(구리포함형) 440mg(25% 수확)을 얻었다.The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water, and immediately adjusted to pH 6.0 with 0.1 N sodium hydroxide. The aqueous solution thus obtained was purified in the same manner as described in step A of Example 1 to obtain 440 mg (25% yield) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 7,985 u / mg. Got it.
[실시예 5]Example 5
NK 631 디하이드로크로라이드(구리포함형)의 합성 메타놀 16ml에 브레오마이신산 3-N-벤조일 아미노프로필에스텔 모노하이드로크로라이드(구리포함형) 9.0g을 용해하였다.Synthesis of NK 631 Dihydrochloride (copper included) 9.0 g of bromycinic acid 3-N-benzoyl aminopropylester monohydrochloride (copper included) was dissolved in 16 ml of methanol.
0℃로 냉각하는 동안 그 용액에 아미노 화합물 9.16g을 가하였다. 0℃에서 40시간 계속 교반한 후 차거운 아세톤을 그 반응 혼합물에 첨가하여 반응생성물을 침전시켰다.9.16 g of amino compound was added to the solution while cooling to 0 ° C. After stirring for 40 hours at 0 ° C., cold acetone was added to the reaction mixture to precipitate the reaction product.
그 침전물을 여과 수집하여 아세톤으로 세척하고 증류수 30ml에 용해시켰다.The precipitate was collected by filtration, washed with acetone and dissolved in 30 ml of distilled water.
그 용액은 0.1N 여산으로 즉시 pH 6.0에 조정하고 0.05M 염화암모니아 수용액에 담긴 CM-Sephadex
염화암모니아 수용액을 세척제로 사용하여 염화암모니아의 농도가 0.05-1.0M까지 계속적으로 증가하는 카람을 통하여 용리제인 용리액 10ℓ를 통과시켜서 정제한다.Ammonia chloride aqueous solution was used as a cleaning agent, and purified by passing 10 liters of eluent, an eluent, through a caram which continuously increased the concentration of ammonia chloride to 0.05-1.0 M.
브레오마이신산에 반응이 안된 3-N-벤조일 아미노프로필에스텔 약 0.2M의 염화암모니아 농도에서 유출물에 나타나고 NK 631은 염화암모니아 농도 약 0.45에서 나타난다.3-N-benzoyl aminopropylester unreacted with bromycin acid appears in the effluent at an ammonium chloride concentration of about 0.2 M and NK 631 is present at an ammonium chloride concentration of about 0.45.
272mμ에서 자외선 흡수를 보이는 그 두 물질을 각기 별도로 수집하였다. NK631를 포함하는 물질은 Amberlite
292mμ에서 자외선 흡수를 보이는 청색물질 총 2.5ℓ를 수집하였다. 증류로서 메타놀을 제거하고 Dowex 44(OH-형)로 pH 6.0에 조정된 후 그 물질을 냉동건조하여 항균효능 8,030u/mg를 갖인 청색 비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 8.2g(90% 수확)을 얻었다. 같은 처리로 미반웅의 브레오마이신산 3-N-벤조일 아미노프로필에스텔 모노하이드로크로라이드(동 포함형) 910mg를 회수하였다.A total of 2.5 L of blue material showing ultraviolet absorption at 292 mM was collected. Ethanol was removed by distillation and adjusted to pH 6.0 with Dowex 44 (OH - type), and then the material was lyophilized to give NK 631 dihydrochloride (formerly copper) in the form of a blue amorphous powder having an antimicrobial effect of 8,030 u / mg. Yield 8.2 g (90% harvest). The same treatment recovered 910 mg of Mibanung Breomycinic Acid 3-N-benzoyl aminopropylester monohydrochloride (included).
[실시예 6]Example 6
NK 631 디하이드로크로라이드(구리포함형) 합성 메타놀 3mℓ에 브레오마이신산 3-N-P-토루엔 설훠넬 아미노프로필 에스텔 모노하이드로크로라이드(구리포함형) 950mg을 용해하였다.NK 631 Dihydrochloride (copper containing) 950 mg of bromycinic acid 3-N-P-toluene sulfonel aminopropyl ester monohydrochloride (copper containing) was dissolved in 3 ml of synthetic methanol.
그 용액에 0℃에서 교반하는 동안 아미노화합물 980mg를 첨가하고 그 혼합물을 0℃에서 48시간동안 교반하였다.To the solution was added 980 mg of the amino compound while stirring at 0 ° C. and the mixture was stirred at 0 ° C. for 48 hours.
그 반응 혼합물에 냉아세톤 15mℓ를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다. 그 침전물을 여과수집하여 아세톤으로 세척하고나서 증류수 30ml에 용해하고 0.1N 염산으로 pH 6.0에 즉시 조정하였다.15 ml of cold acetone was added to the reaction mixture with stirring to precipitate the breomycin component. The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid.
그렇게하여 얻은 용액을 실시예 5의 A 공정에서 설명한 공법에 따라 정제하여 항균효능 8,010u/mg를 갖인 청색 비결정 분말형태의 NK 631 디하이드로크로라이드(구리포함형) 833mg(85% 수확)을 얻었다.The solution thus obtained was purified in accordance with the method described in step A of Example 5 to obtain 833 mg (85% yield) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,010 u / mg. .
[실시예 7]Example 7
NK 631 디하이드로크로라이드(구리포함형)의 합성 디메틸흐름 아마이드 5ml에 브레오마이신산 3-아세틸 아미노프로필 에스텔 모노하이드로크로이드(구리포함형) 850mg을 용해하였다. 그 용액에 0℃에서 교반되는 동안 아미노화합물 950mg를 첨가하고 그 혼합물을 0℃에서 45시간 동안 교반하였다.Synthesis of NK 631 Dihydrochloride (copper containing) 850 mg of breomycinic acid 3-acetyl aminopropyl ester monohydrochromoid (copper containing) was dissolved in 5 ml of dimethyl flow amide. To the solution was added 950 mg of the amino compound while stirring at 0 ° C. and the mixture was stirred at 0 ° C. for 45 hours.
그 반응 혼합물에 차거운 아세톤 20ml를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다.20 ml of cold acetone was added to the reaction mixture with stirring to precipitate the breomycin component.
그 침전물을 여과수집하여 아세톤으로 세척하고나서 증류수 30ml에 용해하여 0.1N 염산으로 pH 6.0에 즉시 조정하였다. 이 용액을 실시예 5의 A 공정와 같은 방법으로 처리하여 항균 효능 8,010u/mg를 갖인 청색비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 570mg(63% 수확)을 얻었다.The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid. The solution was treated in the same manner as in the process A of Example 5, to obtain 570 mg (63% harvest) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,010 u / mg.
[실시예 8]Example 8
NK 631 디하이드로크로라이드(구리포함형)의 합성 디메틸 설프옥사이드 3ml에 브레오마이신산 3-N-(2,4-디니트로훼닐)-아미노프로필 에스텔 모노하이드로크로라이드(구리포함형)를 용해하였다.Synthesis of NK 631 Dihydrochloride (Copper-Containing Type) 3 ml of bromycinic acid 3-N- (2,4-dinitrofenyl) -aminopropyl ester monohydrochloride (copper-containing) was dissolved in 3 ml of dimethyl sulfoxide. .
그 용액에 0℃에서 교반되는 동안 아미노화합물 793mg를 첨가하고 그 혼합물을 0℃에서 24시간 교반하였다.793 mg of the amino compound was added to the solution while stirring at 0 ° C. and the mixture was stirred at 0 ° C. for 24 hours.
그 반응 혼합물에 차거운 아세톤 15ml를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다.15 ml of cold acetone was added to the reaction mixture with stirring to precipitate the breomycin component.
그 침전물을 여과 수집하여 아세톤으로 세척하고나서 증류수 30ml에 용해하여 0.1N 염산으로 pH 6.0에 즉시 조정하였다. 이렇게 된 수용액을 실시예 5의 A 공정에서 설명된 공법에 따라 정제하여 항균효능 8,010u/mg를 갖인 청색 비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 690mg(78% 수확)를 얻었다.The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid. The aqueous solution was purified in accordance with the method described in step A of Example 5 to obtain 690 mg (78% yield) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,010 u / mg. .
[실시예 9]Example 9
NK 631 디하이드로크로라이드(구리포함형) 및 구리비포함 디하이드로크로라이드의 합성 메타놀 10ml에 브레오마이신산 3-아미노프로필 에스텔 디하이드로크로라이드(구리포하형) 1.0g를 용해하였다. 강력하게 교반된 용액에 30분간에 걸쳐 벤조일 크로라이드 104mg를 적하 첨가하였으며 그동안에 그 용액의 pH는 1.4-디아자비싸이크로(2,2,2) 옥탄의 첨가로 6.5에서 7.5까지로 유지하였다. 추가로 30분동안 더 교반한 후 반응 혼합물을 2ml로 농축하였다.Synthesis of NK 631 Dihydrochloride (Copper) and Copper-Free DihydroCride 1.0 g of bromycinic acid 3-aminopropyl ester dihydrochloride (Guripo) was dissolved in 10 ml of methanol. To the vigorously stirred solution was added dropwise 104 mg of benzoyl chloride over 30 minutes, during which the pH of the solution was maintained from 6.5 to 7.5 with the addition of 1.4-diazabicyclo (2,2,2) octane. After a further 30 min stirring the reaction mixture was concentrated to 2 ml.
0℃로 냉각된 농축물에 아미노화합물 1.1g를 교반하면서 첨가하고 그 혼합물을 0℃에서 72시간동안 교반하였다.To the concentrate cooled to 0 ° C., 1.1 g of amino compound was added with stirring and the mixture was stirred at 0 ° C. for 72 hours.
그 반응교환물에 차거운 아세톤 6ml를 교반하면서 첨가하여 브레오마이신 성분을 침전시켰다. 그 침전물을 여과수집하여 아세톤으로 세척하고 증류수 30ml에 용해하여 0.1N 염산으로 pH6.0에 즉시 조정하였다. 이렇게하여 얻은 수용액을 0.05M 염화암모니아 수용액에 담긴 CM-Sephadex
흡착된 물질은 실시예 5의 A공법과 같은 방법으로 처리하여 항균효능 8,000u/mg을 갖인 청색비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 779mg(73% 수확)을 얻었다. 추가하여 미반응의 브레오마이신산 3-N-벤조일 아미노프로필에스텔 모노하이드로크로라이드(구리포함형)을 회수하였다.The adsorbed material was treated in the same manner as in the method A of Example 5 to obtain 779 mg (73% yield) of NK 631 dihydrochloride (copper containing) in the form of blue amorphous powder having an antimicrobial effect of 8,000 u / mg. In addition, unreacted bromycin acid 3-N-benzoyl aminopropylester monohydrochloride (copper included) was recovered.
B공정 증류수 20ml에 위의 A공정에서 얻은 NK 631 디하이드로크로라이드(구리포함형) 700mg을 용해하였다. 그 용액을 증류수에 담긴 Diaion
그리고나서 그 카람을 메타놀과 0.0025N의 염산 수용액(1 : 1V/V)으로 세정하여 290mμ에서 자외선 흡수를 보이는 물질 98ml를 수집하였다.The Karam was then washed with methanol and 0.0025N aqueous hydrochloric acid solution (1: 1V / V) to collect 98 ml of the material showing ultraviolet absorption at 290 mμ.
감압증류로 메타놀을 제거한 후 잔액(殘液)을 Dowex
[실시예 10]Example 10
NK 631 모노설훼이트(구리포함형)와 디아세틱 에시드염(구리비포함형)의 합성Synthesis of NK 631 Monosulfate (Copper Type) and Diacetic Acid Salt (Copper Type)
A공정 : 메타놀 10ml에 브레오마이신산 3-아미노프로필 에스텔 디하이드로크로라이드(구리포함형) 1.0g을 용해하였다. 실내온도에서 강력하게 교반하는 용액에 30분동안 벤조일 크로라이드 104mg을 적하 첨가하였으며 그 동안에 그 용액의 pH는 N-메틸모포린의 첨가로 5.0-7.5에 유지하였다. 30분을 더 교반한후에 그 반응 혼합물에 아세톤 50ml를 첨가하여 브레오마이신 성분을 침전시켰다.Step A: 1.0 g of bromycinic acid 3-aminopropyl ester dihydrochloride was dissolved in 10 ml of methanol. To the vigorously stirred solution at room temperature was added dropwise 104 mg of benzoyl chloride for 30 minutes during which the pH of the solution was maintained at 5.0-7.5 with the addition of N-methylmorpholine. After 30 minutes of further stirring, 50 ml of acetone was added to the reaction mixture to precipitate the breomycin component.
그 침전물을 여과 수집하여 아세톤으로 세척하고 메타놀 3ml에 다시 용해 하였다.The precipitate was collected by filtration, washed with acetone and dissolved again in 3 ml of methanol.
그 메타놀용액에 0℃에서 냉각하고 교반하는 동안에 아미노화합물 597mg을 첨가하였으며 그 혼합물을 0℃에서 42시간 교반하였다.To the methanol solution was added 597 mg of the amino compound while cooling and stirring at 0 ° C., and the mixture was stirred at 0 ° C. for 42 hours.
그 반응 혼합물에 차거운 아세톤 10ml를 교반하면서 첨가하여 반응 생성물을 침전시켰다.10 ml of cold acetone was added to the reaction mixture with stirring to precipitate the reaction product.
그 침전물을 여과수집하여 아세톤으로 세척하고나서 증류수 30ml에 용해하여 0.1N 염산으로 pH를 6.0에 즉시 조정하였다. 그 용액을 실시예 5의 A공정에서와 같은 방법으로 CM-Sephadex
반응 생성물을 포함하는 물질을 증류수에 담긴 Diaion
물로 세척한 후 흡착물을 메타놀과 0.01N 황산수용액(1:1V/V)의 혼합물로 정제하여 292mμ에서 자외선 흡수를 보이는 물질 130ml를 수집하였다.After washing with water, the adsorbate was purified with a mixture of methanol and 0.01 N sulfuric acid solution (1: 1 V / V) to collect 130 ml of a material showing ultraviolet absorption at 292 mμ.
증류로 메타놀을 제거한 후 잔액을 Dowex
위 A공정에서 얻은 NK 631 모노설훼이트(구리포함형) 800mg의 구리제거처리는 실시예 9의 B공정에서와 같은 방법으로 하였다. 단 다른 것은 정제과정에서 5%암모니아 아세테이트 수용액 및 메타놀과 0.01N 초산수용액(1:1V/V)의 혼합물이 각각 5% 염화나트륨 수용액 및 메타놀과 0.0025N 염산수용액(1:1V/V)대신에 사용된 것이다.Copper removal of 800 mg of NK 631 monosulfate (copper-containing type) obtained in step A was performed in the same manner as in step B of Example 9. In addition, a 5% aqueous ammonia acetate solution and a mixture of methanol and 0.01N acetic acid solution (1: 1V / V) were used instead of 5% aqueous sodium chloride solution, methanol and 0.0025N hydrochloric acid solution (1: 1V / V), respectively. It is.
이리하여 엷은 황백색의 비결정부날 상태의 NK 631 디아세틱에서드염(구리포함형) 748mg(96% 수확)을 얻었다. 이 소산물에 자외선 흡수 최대치와 항균효능은 아래에서 보는 바와 같다.This resulted in 748 mg (96% yield) of NK 631 diacetic (copper containing) in pale yellowish white amorphous bun blade. The maximum UV absorption and antimicrobial activity of this product is shown below.
자외선 흡수 최대치UV absorption maximum
NK 631 디하이드로크로라이드(구리포함형) 및 구리비포함디하이드로크로라이드의 합성Synthesis of NK 631 Dihydrochromide (Copper Type) and Copper-Free DihydroCride
메타놀 3ml에 브레오마이신산 3-아미노프로필 에스텔 디하이드로크로라이드 1.0g을 용해하였다.1.0 g of bromycinic acid 3-aminopropyl ester dihydrochloride was dissolved in 3 ml of methanol.
그 용액에 실내온도에서 S-터트-부톡시카보닐-4,6-디메틸-2-멜카프로-피리미딘 193mg을 60분간에 걸쳐 저하시켜 첨가하였다.To the solution was added 193 mg of S-tert-butoxycarbonyl-4,6-dimethyl-2-melcapro-pyrimidine over 60 minutes at room temperature.
그간에 pH는 메타놀에 혼합한 14% 트리에틸아민을 첨가하여 6.0에서 7.4로 유지하였다.Meanwhile, the pH was maintained at 6.0 to 7.4 by adding 14% triethylamine mixed with methanol.
그 혼합물을 브레오마이신산의 3-N-터트-부톡시카보닐 아미노프로필 에스텔이 생성토록 2시간 더 교반하였다. 그리고나서 그 혼합물을 0℃로 냉각하고, 아미노화합물 1.2g를 첨가 혼합하여, 70시간 교반하였다.The mixture was further stirred for 2 hours to form 3-N-tert-butoxycarbonyl aminopropyl ester of bromycinic acid. Then, the mixture was cooled to 0 ° C, 1.2 g of the amino compound was added and mixed, and stirred for 70 hours.
그 반응 혼합물에 차거운 아세톤 10ml을 첨가하여 브레오마이신산을 침전시켰다. 이 침전물을 여과수집하여, 아세톤으로 세척하고 증류수 30ml에 용해하며, 즉시 0.1N 염산으로 pH를 6.0으로 조정하였다.10 ml of cold acetone was added to the reaction mixture to precipitate bromycinic acid. The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water, and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid.
이렇게하여 얻은 수용액을 실시예 5의 A 공정에서 사용한 공법으로 정제하여 항균효능 8,010u/mg를 가진 청색비결정 분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 597mg(56% 수확)을 얻었다.The aqueous solution thus obtained was purified by the method used in the step A of Example 5 to obtain 597 mg (56% yield) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,010 u / mg.
B공정 : 메타놀 50ml에 위의 A공정에서 얻은 NK 631 디하이드로크로라이드(구리포함형) 500mg을 용해하였다. 이 용액에 강력한 교반을 하면서, 내경 1mm의 노즐을 그 용액에 잠기게하여 많은 초과량의 유화수소르르 용해하도록 1시간동안 유화수소를 주입하였다.Process B: In 50 ml of methanol, 500 mg of NK 631 dihydrochromide (copper containing type) obtained in the above process A was dissolved. With vigorous stirring of the solution, hydrogen emulsion was injected for 1 hour to immerse the nozzle with an internal diameter of 1 mm in the solution to dissolve a large amount of excess hydrogen emulsion.
유화수소의 주입완료후, 그 용액을 실내온도에서 30분간 방치하였다. 침전된 유화동을 여과수집하여, 유화수소를 포화시킨 메타놀 50ml로 세척하였다. 여과세척물을 합쳐서, 감압증류로 메타놀과 유화수소로부터 분리하였다.After the completion of the hydrogen emulsion injection, the solution was left at room temperature for 30 minutes. The precipitated copper copper was collected by filtration and washed with 50 ml of methanol, saturated with hydrogen sulfide. The filtrates were combined and separated from methanol and hydrogen emulsion by distillation under reduced pressure.
이 잔액을 메타놀 50ml에 용해하여, 에틸에텔 100ml와 혼합하였다. 형성된 침전물을 여과수집하여 에텔로 세척하고, 건조하여, 7,820u/mg의 항균효능을 갖은 엷은 황백색의 비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 394mg(82% 수확)을 얻었다.This residue was dissolved in 50 ml of methanol and mixed with 100 ml of ethyl ether. The precipitate formed was collected by filtration, washed with ether, and dried to obtain 394 mg (82% yield) of NK 631 dihydrochloride (copper containing) in the form of a pale yellow-white amorphous powder having an antimicrobial effect of 7,820 u / mg.
[실시예 12]Example 12
NK 631 디하이드로크로라이드(구리포함형) 및 구리비포함형의 디하이드로크로라이드의 합성Synthesis of NK 631 Dihydrochromide (Copper Type) and Copper-Free Type DihydroCride
A공정 : 메타놀 2ml에 브레오마이신산 2-아미노프로필 에스텔 디하이드로크로라이드(구리포함형) 1.0g를 용해하였다.Step A: 1.0 g of bromycinic acid 2-aminopropyl ester dihydrochloride was dissolved in 2 ml of methanol.
실내온도에서 강력하게 교반중인 용액에 사리시알데이드 98mg를 트리에틸아민의 첨가로 용액의 pH를 7.0에서 7.4로 유지하면서 첨가하여 브레오마이신산 3-N-사리시리딘아미노프로필 에스텔을 만들게 하였다.To the solution under vigorous stirring at room temperature was added 98 mg of sarisalidate with triethylamine while maintaining the pH of the solution from 7.0 to 7.4 to form 3-brominemyriate 3-N-sarishiridineaminopropyl ester.
한시간의 계속된 교반후, 0℃로 냉각한 그 혼합물에 아미노 화합물 600mg을 교반하면서 첨가하였다.After one hour of continued stirring, 600 mg of the amino compound was added to the mixture which was cooled to 0 ° C. with stirring.
이 혼합물을 30시간동안 교반하였다. 그 반응 혼합물에 차거운 아세톤 10ml를 첨가하여 브레마이신산을 침전시켰다. 침전물을 여과수집하여, 아세톤으로 세척하고나서 증류수 30ml에 용해하여 즉시 0.1N 염산으로 pH를 6.0에 조정하였다. 이렇게 얻은 수용액을 실시예 5의 A공정에서와 같이 정제하여 8,013u/mg의 항균효능을 갖인 NK 631 디하이드로크로라이드(구리포함형) 501mg(47% 수확)을 얻었다.The mixture was stirred for 30 hours. 10 ml of cold acetone was added to the reaction mixture to precipitate bremic acid. The precipitate was collected by filtration, washed with acetone, dissolved in 30 ml of distilled water and immediately adjusted to pH 6.0 with 0.1 N hydrochloric acid. The aqueous solution thus obtained was purified as in the step A of Example 5 to obtain 501 mg (47% harvest) of NK 631 dihydrochloride (copper containing type) having an antibacterial effect of 8,013 u / mg.
B공정 : 0.5N 염산 수용액 25ml에 위 공정에서 얻은 NK 631 디하이드로크로라이드(구리포함형)을 150mg용 해하였다. 그 용액에 0.2%의 디치존(디페닐치오카바존)을 포함한 크로로홈용액 25ml를 첨가하였다Process B: 150 mg of NK 631 dihydrochloride (copper included) obtained in the above process was dissolved in 25 ml of 0.5N hydrochloric acid aqueous solution. To the solution was added 25 ml of crorohome solution containing 0.2% of dizone (diphenylthiocarbazone).
급속교반 및 혼합후에 그 혼합물을 정치하도록 방치하여 2개의 층으로 분리하게 하였다.After rapid stirring and mixing the mixture was left to stand to separate into two layers.
아래쪽층(크로로홈층)은 뽑아내고, 윗층부를 디치존을 포함한 새크로로홈 용액 25ml와 혼합하였다.The lower layer (chromo groove layer) was removed and the upper layer was mixed with 25 ml of sacro groove solution including dichzone.
위의 교반, 층계분리 및 디치존을 포함하는 새로운 크로로홈 용액의 첨가등의 공정을 8회 반복하여 마지막에 윗층부(수성층)을 크로로홈으로 세척하고, 분리한 수성층을 Dowex 44(OH-형)으로 pH6.0에 조정한다.Repeat the above steps 8 times, such as stirring, stair separation, and addition of a new crohome solution containing dichzone, and finally, the upper layer (aqueous layer) was washed with crow groove, and the separated aqueous layer was washed with Dowex 44 (OH - the type) is adjusted to pH6.0.
이렇게 처리된 수성층을 건조될때까지 감압하에 증발시켜, 7,845u/mg의 항균효능을 갖인 엷은 황백색의 비결정분말 형태의 NK 631 디하이드로크로라이드(구리포함형) 415mg(96% 수확)을 얻었다.The treated aqueous layer was evaporated under reduced pressure to dryness to obtain 415 mg (96% yield) of NK 631 dihydrochloride (copper containing type) in the form of a pale yellow-white amorphous powder having an antimicrobial effect of 7,845 u / mg.
[실시예 13]Example 13
NK 631 디하이드로크로라이드(구리포함형)의 합성Synthesis of NK 631 Dihydrochloride (Copper Type)
메타놀 2ml에 브레오마이신산 3-아미노프로필 에스텔 디하이드로 크로라이드(구리포함형) 1.0g을 용해하였다.1.0 g of bromycinic acid 3-aminopropyl ester dihydro chromide (copper containing type) was dissolved in 2 ml of methanol.
0℃로 냉각한 용액에 아미노화합물 1.0g을 교반하며 첨가하고 그 혼합물을 72시간동안 더 교반하였다.1.0 g of the amino compound was added to the solution cooled to 0 ° C. with stirring, and the mixture was further stirred for 72 hours.
반응혼합물에 차거운 아세톤 6ml를 교반하며 첨가하여 브레오마이신산을 침전시켰다.6 ml of cold acetone was added to the reaction mixture with stirring to precipitate bromycinic acid.
그 침전물을 실시예 5의 A공정에서와 같이 정제하여, 8,015u/mg의 항균효능을 보유한 청색비결정분말형태의 NK 631 디하이드로크로라이드(구리포함형) 128mg(12% 수확)을 얻었다.The precipitate was purified as in step A of Example 5 to obtain 128 mg (12% harvest) of NK 631 dihydrochloride (copper containing) in the form of a blue amorphous powder having an antimicrobial effect of 8,015 u / mg.
[참고예 1]Reference Example 1
N-[(S)-1'-페닐에틸]-1,3-디아미노푸로반의 합성Synthesis of N-[(S) -1'-phenylethyl] -1,3-diaminofuroban
0℃냉각 상태에서 (S)-1-페닐에틸아민 50g에 아코리로니트릴 33g을 교반하면서 첨가하였다.33 g of acrylonitrile was added to 50 g of (S) -1-phenylethylamine while stirring at 0 ° C.
이 혼합물을 염화칼슘에 연결된 리후럭스콘덴서가 달린 후라스크에 담고 93℃로 가열하며, 그 온도로 18시간동안 교반되게 한다. 반응 완료후, 반응혼합물을 감압증류로 초과 아크리토니트릴로부터 분리한다.The mixture is placed in a flask with a reflux condenser connected to calcium chloride and heated to 93 ° C. and allowed to stir at that temperature for 18 hours. After completion of the reaction, the reaction mixture is separated from excess acritonitrile by distillation under reduced pressure.
그 잔액을 감압증류하고 140℃ 내지 145℃/7mmHg에 끓는 한 물질을 수집하여 3-[(S)-1'-페닐에틸아미노]프로피오니트릴 53.5g을 얻는다.The residue was distilled under reduced pressure and one substance was collected at 140 ° C. to 145 ° C./7 mmHg to collect 53.5 g of 3-[(S) -1′-phenylethylamino] propionitrile.
이 니트릴을 레이니 닛켈 W-7과 암모니아 15%를 포함한 에타놀 50ml와 함께 고압반응관에 넣는다.The nitrile is placed in a high pressure reactor tube with 50 ml of ethanol containing Raney Nikel W-7 and 15% ammonia.
그 혼합물을 50°내지 58℃와 수소부분압력 100 내지 40kg/cm2에서 1.5시간동안 1,000rpm의 속도로 교반하였다. 분해 완료후, 반응혼합물을 감압으로 증류하였다.The mixture was stirred at 50 rpm to 58 캜 and hydrogen partial pressure at 100 to 40 kg / cm 2 for 1.5 hours at a speed of 1,000 rpm. After completion of the decomposition, the reaction mixture was distilled off under reduced pressure.
95°-103℃/2mmHg에서 끓는 한 물질을 수집하여 N-[(S)-1'-페닐에틸]-1,3-디아미노푸로판 48.3g을 얻었다.One boiling material was collected at 95 ° -103 ° C./2 mmHg to obtain 48.3 g of N-[(S) -1′-phenylethyl] -1,3-diaminofurophane.
[(S)-1-페닐에틸아민으로부터의 이론산출량의 66%][66% of Theoretical Output From (S) -1-phenylethylamine]
Claims (1)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR7801991A KR810001407B1 (en) | 1978-06-28 | 1978-06-28 | Preparation of 3-[(S) -1′-phenylethyl amino] propyl amino bromycin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR7801991A KR810001407B1 (en) | 1978-06-28 | 1978-06-28 | Preparation of 3-[(S) -1′-phenylethyl amino] propyl amino bromycin |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR810001407B1 true KR810001407B1 (en) | 1981-10-20 |
Family
ID=19208060
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR7801991A Expired KR810001407B1 (en) | 1978-06-28 | 1978-06-28 | Preparation of 3-[(S) -1′-phenylethyl amino] propyl amino bromycin |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR810001407B1 (en) |
-
1978
- 1978-06-28 KR KR7801991A patent/KR810001407B1/en not_active Expired
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