CN1102853C - Tumor metastasis-suppressing agent and veterinary tumor metastasis-suppressing agent - Google Patents
Tumor metastasis-suppressing agent and veterinary tumor metastasis-suppressing agent Download PDFInfo
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- CN1102853C CN1102853C CN93115007A CN93115007A CN1102853C CN 1102853 C CN1102853 C CN 1102853C CN 93115007 A CN93115007 A CN 93115007A CN 93115007 A CN93115007 A CN 93115007A CN 1102853 C CN1102853 C CN 1102853C
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Abstract
The present invention provides a carcinomatosis inhibitor and a carcinomatosis inhibitor for animals, which is characterized in that the inhibitors contain polypeptides represented by the amino acid sequences or the mixture of the polypeptides: X-X'-(the amino acid sequence of the fourth exterior contact part of TNF-alpha), wherein V is hydrogen atoms or random kinds and number of peptides, and X' is peptides of which the amino acid residues are from 1 to 39; but except that X-X' is Met-Val-Arg-Ser-Ser-Ser-Arg-The-Pro-Ser-Asp-Lys-Pro-Val-Ala-His-Val-Val.
Description
The present invention relates to cancer transfer inhibitor and veterinary tumor metastasis-suppressing agent.
Tumor is the spontaneous superfluous proliferating cells group of energy.Though can distinguish the malignant tumor (or being called cancer) and the benign tumor different that cause the animal dead probability with them, many exceptions are also arranged, it is inconvenient (" rock ripple diction biology is bent " the 3rd edition page 4) that both strictnesses are made a distinction
At present, the treatment method for cancer has three kinds, i.e. surgical treatment, pharmacotherapy and actinotherapy.What in fact extensively adopt is with above-mentioned therapy, perhaps and then be the Therapeutic Method that above-mentioned therapy and laser therapy etc. are combined.But consider the misery in when treatment and the transfer of cancer, that yes is very urgent for the expectation of pharmacotherapy, therefore should study and use anticarcinogen in large quantities.Yet be effectively without any a kind of anticarcinogen to various cancers also, simultaneously because use can not be given full play to effect separately, and the effect of paying is big, and majority can not life-time service, the therefore general therapy that adopts various medicaments and usefulness.Therefore very the anticarcinogen, particularly anticancer effect that make new advances of expectation development is good, easy and the medicament that can life-time service of method.
In addition, cancer has the feature that can shift.The reason of cancer patient's death almost is because cancerometastasis entirely.Its reason of saying of so-called " controlled to shift and just controlled cancer " just is this.But present anticarcinogen all is that anticancer effect with the cancer cell multiplication of former position that suppresses cancer and metastasis site is a target, yet there are no to have the report that suppresses the cancerometastasis effect in the anticarcinogen that uses at present.In addition, it is reported, can promote the transfer of cancer as peptide T NF-α with anti-tumor activity.(PeterOrosz etc. " enhancement of Experimental Metastasis by Tumor Necro-sis Factor ", J.Exp.Med., Vol.177, pp 1391-98,1993.5, The Rockefeller University Press).In sum, can think that anticancer effect and cancerometastasis suppress effect and be based on the different mechanism of action.
To suppress cancerometastasis is that the exploitation of the medicament of target still is few.Though also reported the sulfated polysaccharide class, N-diazonium acetoglycocoll derivant, the sour sweet enzyme of neural ammonia (sugar) (above-mentioned for growing work " sending out and shifting of cancer " 60-70 page or leaf,, society of free the Republic of China in 1981 in the island) fiber is in conjunction with catabolic enzyme (FNS), (above-mentioned be to compile " cancer dictionary " with Tian Wuxiong to people's r TIMP, 82 pages, nineteen ninety, Japan comment society) inhibited, but the report of practicability is not arranged as yet, do not have them itself to have the report of antitumaous effect yet.
Consider the importance of transfer inhibitor aspect the control cancer, we can say that the utmost point expects that urgently exploitation has above-mentioned inhibiting medicament.
Technology contents of the present invention provides novel cancer transfer inhibitor and novel veterinary tumor metastasis-suppressing agent.
The inventor provides and has contained the polypeptide that useful following aminoacid sequence represents or the cancer transfer inhibitor of its mixture, thereby has finished the present invention.X-X’-Ala-Asn-Pro-Gln-Ala-Glu-Gly-Gln-Leu-Gln-Trp-Leu-Asn-Arg-Arg-Ala-Asn-Ala-Leu-Leu-Ala-Asn-Gly-Val-Glu-Leu-Arg-Asp-ASn-Gln-Leu-Val-Val-Pro-Ser-Glu-Gly-Leu-Tyr-Leu-Ile-TyrSer-Gln-Val-Leu-Phe-Lys-Gly-Gln-Gly-Gln-Gly-Cys-Pro-Ser-Thr-His-Val-Leu-Leu-Thr-His-Pro-Ser-Thr-His-Val-Leu-Leu-Thr-His-Thr-Ile-Ser-Arg-Ile-Ala-Val-Ser-Tyr-Gln-Thr-Lys-Val-Asn-Leu-Leu-Ser-Ala-Ile-Lys-Ser-Pro-Cys-Gln-Arg-Glu-Thr-Pro-Glu-Gly-Ala-Glu-Ala-Lys-Pro-Trp-Tyr-Glu-Pro-Ile-Tyr-Leu-Gly-Gly-Val-Phe-Gln-Leu-Gln-Lys-Gly-Asp-Arg-Leu-Ser-Ala-Glu-Ile-Asn-Arg-Pro-Asp-Tyr-Leu-Asp-Phe-Ala-Glu-Ser-G1y-Gln-Val-Tyr-Phe-Gly-Ile-Ile-Ala-Leu。(X is that the peptide X ' of hydrogen atom or any kind, quantity is the peptide of 1-39 amino acid residue)
Above-mentioned aminoacid sequence, promptly connect X ' from Ala Leu to the end, the aminoacid sequence of the 4th circumscribed (エ Network ソ Application) part that has been connected the TNF-α of guanine with known head at base sequence is identical.Therefore, in the other parts of this description, above-mentioned aminoacid sequence is represented with X-X '-(aminoacid sequence of the 4th circumscribed part of TNF).
Aforementioned polypeptides and antitumaous effect thereof are that the applicant speciallys permit out the clear 61-169522 of Willing number content in Japan, and are disclosed in open special permission communique according to 62-282587 number.
Cancer transfer inhibitor of the present invention and veterinary tumor metastasis-suppressing agent contain polypeptide that the above-mentioned aminoacid sequence of a kind of usefulness represents or the mixture more than 2 kinds that contains them.And then the dosage form needs according to preparation suitably cooperate excipient, disintegrating agent, and drug flavorings etc. can be prepared into as lozenge, powder, various dosage forms such as injection.Only otherwise its cancerometastasis of overslaugh suppresses active, preparation of the present invention does not limit and uses specific auxiliary agent and specific dosage form.
For the cancerometastasis that confirms cancer transfer inhibitor of the present invention and veterinary tumor metastasis-suppressing agent suppresses effect, to mice body angular vein injection cancerous cell, with the naked eye number goes out the cancerous node number of its pulmonary.In the case, though different owing to the difference at former position of cancer, in general, in blood transfer as the cancerometastasis mode, lymph metastasis, during destructive transfer and contact shifted, blood transfer and lymph metastasis were maximum.Passing through under the situation of lymph metastasis, cancerous cell finally enters blood through lymphatic vessel.Simultaneously and since to lung shift the easiest observed (the rich work of aforesaid " sending out again and shifting of cancer " the 6th page and holt " oncology " the 40th page, 1984, Nan Shantang), therefore can think and observe that to shift to pulmonary by blood be only.
The dosage of cancer transfer inhibitor of the present invention and veterinary tumor metastasis-suppressing agent, dosing interval, should be under doctor and veterinary's strictness be instructed, according to the age of dispensing object, symptom, body weight, the dispensing effect determines respectively, but for adult (60kg), every day, dispensing standard roughly was: oral administration medicine supplying 1 * 10
7Unit, the vein dispensing is 1 * 10
6Unit, subcutaneous administration is 5 * 10
5Unit.Can once-a-day or divide dispensing for several times.Also have, for large animals such as the oxen and horses, the dosage of per kilogram of body weight is 1/60 of an above-mentioned throwing amount, for pig, and dog, toy in retouching etc., per kilogram of body weight is thrown 2 times of above-mentioned amount, is 2 times of middle toy for the dispensing standard of birds pers kilogram of body weight such as chicken.
Above-mentioned unit is the index of Asahi Kasei Corporation of manufacturer in order to the concentration of expression TNF-α.Is benchmark with various peptides for the cytotoxicity of L-929 cell [Proceeding of national a-cademy science ofusA 72,3666-3670 page or leaf], and available following method is obtained.
Cytotoxicity to the L929 cell
With the L929 cell, in being added with the Eagles mininumessential culture medium (hereinafter referred to as the MEM culture medium) of 5% foetal calf serum, cultivate, will contain 8 * 10 in the above-mentioned culture medium of 100 μ l
4The sample of individual cell is put in the 96 cave plates and cultivates, and condition of culture is 37 ℃, and 4-6 hour, 5%CO
2, 100% water, and the method for the cultured cell of use routine.Add D actinomycin D then in culture medium, making its ultimate density is 1 μ g/ml, and this moment, the amount of culture fluid was 150 μ l.(so D actinomycin D is the medicament that often uses because of improving cellular sensitivity, and itself is to L929 cell avirulence).Add the body that tried immediately, (suitably adjust dilution rate this moment so that can obtain ED with the suitable dilution of 50 μ l MEM culture medium
50Value), be that the L929 cell of 200 μ l was cultivated 18 hours under above-mentioned condition with final liquid measure.For measuring the necrocytosis activity, at first remove culture medium, add the fixing dyeing of 2% methanol solution that contains 0.2% crystal violet again, because crystal violet can make nucleated cell dyeing, because the result of necrocytosis, can not be colored by the bottom surface of the ware cell that dissociates, therefore can directly measure the necrocytosis activity.Be used for OD
590nmThe absorption measurement dyeability at place and the dyeability comparison of matched group just can be measured the necrocytosis activity, active being defined as follows.
Obtain the dilution rate of being tried body (N) that can make L929 cells survival 50%, use rabbit TNS to compare, with 2.4 * 10
4The TNF α of unit/mg/ml determines this to exempt from the active n of TNS (unit/ml), obtain rabbit TNS ED
50Dilution rate (C).
Tried body activity (unit/ml) calculate with N/c * n.
Wear in the row at above-mentioned aminoacid, X-X ' is the polypeptide (hereinafter referred to as TNF-AM1) of Met-Val-Arg-Ser-Ser-Thr-Arg-Thr-Pro-Ser-Arg-Lys-Pro-Val-Ala-His-Val-Val, and it is to the LD of Mus
50Value is:
9.5 * 10
7Unit/kg (BALB/c system)
2.7 * 10
7Unit/kg (C3H/He system)
5.8 * 10
7Unit (C57BL/6 system); X-X ' is: the polypeptide of Met-Val-Arg-Ser-Cys-Thr-Arg-Thr-Pro-Ser-Arg-Lys-Pro-Val-Ala-His-Val-Val, (hereinafter referred to as TNF-AM2), it is to the LD of Mus
50Value is: 2.9 * 10
7Unit/kg (BALB/c system)
2.7 * 10
7Unit/kg (C3H/He system)
2.7 * 10
7Unit/kg (C57BL/6 system) all is values of TNF-α, with
1.0 * 10
7Unit/kg (BALB/c system)
7.8 * 10
6Unit/kg (C3H/He system)
1.5 * 10
7Unit/kg (C57BL/6 system) compares much bigger, therefore as above-mentioned illustrated, polypeptide of the present invention is for Normocellular toxicity, and is more much lower than TNF-α, and cancer transfer inhibitor of the present invention and veterinary tumor metastasis-suppressing agent are extremely useful clinically.
With following embodiment, experimental example explains the present invention.
Embodiment 1 (injection)
With 1 * 10
6The TNF-AM1 of unit, or TNF-AM2 or their mixture be dissolved in the normal saline solution, is mixed with intravenous fluid.
Embodiment 2 (slow releasing agent)
With 3 * 10
6The TNF-AM1 of unit, or in TNF-AM2 or dispersion of their mixture and the inclosure ribosome, make slow releasing agent.
Embodiment 3 (tablet)
TNF-AM2 1g
6%HPC lactose 178g
Stearic acid Pulvis Talci 8g
Potato starch 14g
Above-mentioned substance is mixed, and tabletting is made 400 in the tablet of the 0.5g of 2.5mg TNF-AM2.
Embodiment 4 (mixture for internal use)
TNF-AM2 2.5g
Pure water 100ml
Embodiment 5 (ointment)
TNF-AM2 0.5g
Refined wool fat 80g
Yellow vaseline is an amount of
1000g experimental example 1
Inject Mus B16 melanotic tumor cell (5 * 10 from the tail vein in the body to mice (C57BL/6, male, average 8 ages in week, average weight 25g)
4Individual), and, throw trial drug after 6,10 days in 2,4, inject the melanotic tumor cell and cuts the taking-up lung after 14 days, the cancerous node number of the pulmonary that detects by an unaided eye, experiment condition and the results are shown in table 1.
Underproof medicine is TNF-α (4.9 * 10
7Unit/mg) and TNT-AM2 (9 * 10
7Unit/mg), all do not observe the dead example of Mus.
Table 1
The average cancer knot of dosage dispensing group number of mice
(unit/time/only) a joint number/A) nothing 6 24.7 ± 10.4B) TNF-α 5 14.7 * 10
489.8 ± 27.1C) 4 4.9 * 10
472.5 ± 69.6D) TNF-AM2 6 30.0 * 10
414.0 ± 5.7E) 6 9.0 * 10
419.5 ± 6.9
The available Fig. 1 of the result of above-mentioned table 1 represents that Fig. 1 clearly illustrates that according to TNF-α consumption and promoted cancerometastasis that TNF-AM2 of the present invention compares with the group of not offeing medicine, and has suppressed 1/2 cancerometastasis.In addition, among the figure A-E be exactly the table in the dispensing group.Experimental example 2
In order to confirm the preventive effect of cancer transfer inhibitor of the present invention, dispensing and observe it before injecting cancerous cell.
Interior to every group of 8 Mus (BALB/c, average 8 ages in week, average weight 25g) body with 1 * 10
3Unit/only, throw TNF-AM2 of the present invention for the F group, injects the solid type cancer Meth A meat tumor cell of Mus through the tail vein simultaneously, for the G group, injects above-mentioned tumor cell after 3 hours and (is 1 * 10
6Individual/only), injects and cuts and take out lung after 21 days, the cancerous node number of the pulmonary that detects by an unaided eye, the result is 83.5 ± 51.4 cancerous nodes of F group average out to, and the G group to be reduced to 1/8 be 10.1 ± 10.7 cancerous nodes.This explanation, it is very important throwing cancer transfer inhibitor of the present invention in advance, that is to say, takes cancer transfer inhibitor of the present invention after the cancer treatment at former position, might suppress sending out again of cancer.
The invention provides novel cancer transfer inhibitor and veterinary tumor metastasis-suppressing agent.
Cancer transfer inhibitor of the present invention and veterinary tumor metastasis-suppressing agent have anticancer effect, and are extremely effective for suppressing sending out again of cancer.Simultaneously low for Normocellular toxicity, but life-time service is very high as cancer transfer inhibitor and its value of veterinary tumor metastasis-suppressing agent.
Fig. 1 represents that cancerometastasis suppresses effect.
Symbol description: not dispensing group of A, B, C throw the group with TNF-α, and D, E throw the group with TNF-AM2.
Claims (2)
1. the cancer transfer inhibitor that contains the polypeptide that useful following aminoacid sequence represents,
X-X '-(aminoacid sequence of the 4th circumscribed part of TNF-α)
X is the peptide of 1 hydrogen atom or any kind, quantity, and X ' is that total number of atnino acid is 1-39 a peptide,
But X-X ' is
Except when Met-Val-Arg-Ser-Ser-Ser-Arg-Thr-Pro-Ser-Asp-Lys-Pro-Val-Ala-His-Val-Val-or Ser-Ser-Ser-Arg-Thr-Pro-Ser-Asp-Lys-Pro-Val-Ala-His-Val-Val-.
2. the veterinary tumor metastasis-suppressing agent that contains the polypeptide of representing with the aminoacid sequence of claim 1 record.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3357352A JPH06263650A (en) | 1991-12-02 | 1991-12-02 | Lps-containing antitumor agent and antitumor agent for animal |
| CN93115007A CN1102853C (en) | 1991-12-02 | 1993-10-15 | Tumor metastasis-suppressing agent and veterinary tumor metastasis-suppressing agent |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP3357352A JPH06263650A (en) | 1991-12-02 | 1991-12-02 | Lps-containing antitumor agent and antitumor agent for animal |
| CN93115007A CN1102853C (en) | 1991-12-02 | 1993-10-15 | Tumor metastasis-suppressing agent and veterinary tumor metastasis-suppressing agent |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1101572A CN1101572A (en) | 1995-04-19 |
| CN1102853C true CN1102853C (en) | 2003-03-12 |
Family
ID=36928374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN93115007A Expired - Fee Related CN1102853C (en) | 1991-12-02 | 1993-10-15 | Tumor metastasis-suppressing agent and veterinary tumor metastasis-suppressing agent |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JPH06263650A (en) |
| CN (1) | CN1102853C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1961823B1 (en) * | 2005-11-28 | 2024-06-12 | Biomedical Research Group Inc. | Method for producing lipopolysaccharide and lipopolysaccharide |
| CN101490092B (en) | 2006-07-14 | 2011-11-09 | 生物医学研究集团有限公司 | Method for production of limulus-positive glycolipid, limulus-positive glycolipid, and limulus-positive glycolipid blend |
| JP6833418B2 (en) * | 2016-09-12 | 2021-02-24 | 有限会社バイオメディカルリサーチグループ | Lipopolysaccharide, lipopolysaccharide production method and lipopolysaccharide formulation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0477791A2 (en) * | 1990-09-21 | 1992-04-01 | Ishihara Sangyo Kaisha, Ltd. | Polypeptide |
-
1991
- 1991-12-02 JP JP3357352A patent/JPH06263650A/en active Pending
-
1993
- 1993-10-15 CN CN93115007A patent/CN1102853C/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0477791A2 (en) * | 1990-09-21 | 1992-04-01 | Ishihara Sangyo Kaisha, Ltd. | Polypeptide |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1101572A (en) | 1995-04-19 |
| JPH06263650A (en) | 1994-09-20 |
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