CN103721245B

CN103721245B - Thymopeptide application in the reagent preparing reversing tumor associated gene mutation

Info

Publication number: CN103721245B
Application number: CN201410003120.7A
Authority: CN
Inventors: 王弢; 渠香云; 秦勇
Original assignee: SUZHOU MICRO DIAG BIOMEDICINE CO Ltd
Current assignee: Jiangsu Microdiag Biomedicine Technology Co ltd
Priority date: 2014-01-03
Filing date: 2014-01-03
Publication date: 2016-08-17
Anticipated expiration: 2034-01-03
Also published as: CN103721245A

Abstract

The invention discloses Thymopeptide application in the reagent preparing reversing tumor associated gene mutation, widen the range of application of Thymopeptide, tumor patient can make tumour associated gene mutation reverse after intervening Thymopeptide, reduce the Sudden Changing Rate of tumor-related gene, hypertrophy or polyp reduce, extend the life cycle of tumor patient, for the thinking that the targeted therapy offer of tumor is new, there is important clinical meaning.

Description

Thymopeptide application in the reagent preparing reversing tumor associated gene mutation

Technical field

The invention belongs to chemical field, particularly to Thymopeptide for preparing the examination of reversing tumor associated gene mutation Application in agent.

Background technology

Cancer is the result of gene mutation accumulation.In other words, the generation of all cancers, it is derived from DNA (deoxyribonucleic acid) (DNA) exception of sequence.International cancer genome co-plan participates in one of unit, univ cambridge uk's Mulberry lattice institute seat of honour Scientist Jake Stratton professor points out, the formation of canceration is the most longer through 8-10, and this very long time is exactly in fact One polygenic mutation the process accumulated.Tumor-related gene can be divided into proto-oncogene and antioncogene, wherein plays main work Sudden change belong to driving sudden change (driver mutation).When Tumor Suppressor Gene Mutations loses suppression cancer effect, or former cancer Gene mutation activates, and will induce cancer.At normal cell during cancerous cell transformation, there is multiple difference former cancer base Cause and Tumor Suppressor Gene Mutations.Comparing conventional use of oncoprotein blood serum designated object in Clinical detection, sudden change is the most special Tumor marker.In December, 2009, Mulberry lattice institute publishes the article declaration on " naturally " magazine, and they take the lead in decoding in the world lung 5 kinds of tumor full gene passwords such as cancer, skin carcinoma and breast carcinoma, draw out corresponding oncogene mutation map.

Some conventional researchs show to involve, in tumor tissues, peripheral blood (plasma/serum), tumor, the body fluid that organ is relevant All it appeared that the mutant DNA cancerous tumor cell that tumor is correlated with can be with released dna to peripheral blood or body fluid in (such as saliva, expectorant etc.) In, thus the sudden change of tumor-related gene can be detected.

Lung cancer related gene has EGFR, KRAS etc., detection EGFR, KRAS gene mutation can find pulmonary carcinoma extreme early signal, Understand the development prognosis of pulmonary carcinoma, assessment chemicotherapy effect etc.；Breast carcinoma related mutation gene BRCA1, BRCA2, RAD51C etc.；Intestinal Cancer related mutation Gene A PC, KRAS, PTEN, TP53, BRAF etc..EGFR exons 19 is pulmonary carcinoma common mutations, and document report is prominent Control with changed scale is 30-40%；Apc gene exons 15 is intestinal cancer common mutations site, and document report mutant proportion is 70-80%； BRCA2 is breast carcinoma common mutations site, document report mutant proportion 40-50%.It addition, molecular targeted agents is according to tumor Cell and normal cell molecular biological characteristics difference and research and develop, such as Iressa (Iressa), Arastin (Avastin) Deng, for blocking the new type anticancer medicine of tumor cell signal transduction pathway, but they only have special gene sudden change or swollen to part The patient that tumor markers is expressed has preferable curative effect.

Up to the present the mankind the most do not break through the difficulty of oncotherapy, although chemotherapeutics and radiotherapy are to tumor cell Lethal effect is affirmative, but Normocellular destruction also be can not be ignored by it.Development along with scientific research And progress, immunomodulator have also been obtained good development and application.In the therapeutic process of tumor, use immunomodulator can With the immunocompetence of enhancing body, impaired medullary cell and immunocyte during protection chemotherapy and radiation.Thymopeptide Being a kind of widely used immunomodulator in clinical therapy of tumor, its Main Function is to promote T cell differentiation, maturation, energy Induction pre T lymphocyte is converted into T cell, and is divided into the T cell subgroups such as Th, Ts, Tc further；Mature T cells antagonism can be strengthened Former or other stimulate reactions, as strengthen PHA or ConA induction lymphproliferation response, promote interleukin-2 generate, increase Strong immunity rejection and graft versus host disease, improve the activity of NK cell, and can strengthen superoxide dismutase in serum Activity.Zoopery shows, it can make thymic mouse part or close to recovering fully, and the lymphoid tissue that can make atrophy is multiple Raw, lymphopoiesis, it is that prolymphocyte is ripe, becomes the lymphocyte having immunologic function.Thymopeptide is controlled in tumor Application in treatment mainly has thymosin, Thymosin alpha 1, Thymopentin three class.

Thymosin is a kind of many peptides preparation extracted from newborn calf thymus tissue, and main chemical compositions is TF5, tool There is regulation and strengthen the effect of human body cell immunologic function, T lymphocyte maturation can be promoted.The report 66 example pulmonary carcinoma such as Deng Qin is suffered from Person, random packet analysis routine blood test and immunologic function, result thymosin group indices is all significantly improved than before treatment.Cheng Rui Tinkling of pieces of jade report patients with gastric cancer after curative dissection 62 example divides two groups of observations, and there were significant differences for Effect of Thymus Peptide Therapy result.The retrospective spy such as Huang Ling Begging for 63 example low doses uses thymosin on Old patients with tumor chemical therapy toxic side effect and the impact of quality of life, have also been obtained certainly Curative effect.In sum, thymosin has promotion cells in vivo cytokine secretion and the effect of lymphocyte function, can enhancing body Immunologic function, oncotherapy plays important assosting effect.

Thymosin alpha 1 can stimulate peripheral blood lymphocyte mitogen, thus promotes the maturation of T lymphocyte, increases antigen Or interferon-ALPHA, interferon gamma and the lymphokine level such as interleukin-2, interleukin-3 that after mitogen activation, T cell is secreted, Increase T cell surface lymphokine receptor level simultaneously.It is also by the activation to cd4 cell, strengthens allosome and autologous Mixed lymphocyte reaction.Set sail etc. to 80 example postoperative NSCLC patient random packet, observe NSCLC postoperative patient chemotherapy combined The recent reaction of Thymosin alpha 1, the every immune indexes of result and numeration of leukocyte significantly improve.Li Yanli is by looking back constitutional Hepatocarcinoma 37 example random packet is observed, and finds that Thymosin alpha 1 can obviously improve immunologic function.Xu Ran etc. by 14 examples through Thymosin alpha The Pancreas cancer patients immune indexes of 1 treatment detects, and compares with the patient not using the same period Thymosin alpha 1 to treat, also Obtain same result.Many report display Thymosin alpha 1s are a kind of biological respinse regulatory factors, it can promote T cell and from The differentiation of Natural killer cell (NK) the change effect affirmative with ripe, to tumor patient immunologic function.

Thymopentin is immunity two-ways regulation agent, has induction and promotes T lymphocyte and subpopulations, maturation and live The function changed, the ratio of regulation T lymphocyte, make CD4/CD8 tend to normal；Regulation and the work of enhancing human body cell immunologic function With, the T lymphocyte maturation in peripheral blood after mitogen can be promoted to activate, increasing T cell has silk at various antigens or cause After mitogen activation, the secretion of various lymphokines, increases lymphokine receptor level in T cell；It is simultaneously by assisting T The activation of cell strengthens lymphocyte reaction, the most also can strengthen NK cytotoxicity.Lin Fang is by NSCLC patient 200 example Random packet carries out comparative study, evaluates Thymopentin effect in NSCLC, finds that Thymopentin treatment group patient's immunity refers to Mark and numeration of leukocyte significantly improve.Thymopentin (TP-5) is the effective ingredient of thymic neuroendocrine carcinoma, by arginine, relies ammonia Acid, aspartic acid, valine, tyrosinase 15 kind aminoacid composition, have identical physiological function and drug effect with thymosin, its Feature is that pharmaceutical purity is high, stable content (for activating the optimal dose of T-lymphocyte), safe and reliable, and do not contain macromole Protein, its effective ingredient is animal thymus extract 84-102 times, and TP-5 has makes superoxide dismutase in blood (SOD) Content raises, the function being remarkably decreased towards Atomic oxygen radical anion content.

The studies above show Thymopeptide as a kind of immunomodulator, the effect in oncotherapy is affirmative, But Thymopeptide reversing tumor extreme early suddenlys change, improves the survival of patients of tumor post-operation poor prognosis, oncotherapy effect, tumor Effect in terms of recurrence monitoring has no report.

Summary of the invention

In view of this, it is an object of the invention to provide Thymopeptide for preparing reversing tumor associated gene mutation Application in reagent.

For achieving the above object, it is provided that following technical scheme:

Thymopeptide application in the reagent preparing reversing tumor associated gene mutation.

Preferably, sudden change in early days, curative effect instruction sudden change, prognosis instruction sudden change or recurrence monitoring sudden change are sported described in.

Preferably, described tumor-related because of proto-oncogene or antioncogene.

It is furthermore preferred that described proto-oncogene is EGFR, KRAS, C-KIT or PIK3CA.

It is furthermore preferred that described antioncogene is P53, APC, BRCA1, BRCA2, PTEN or NOEY2.

Most preferably, described Thymopeptide is thymosin, Thymosin alpha 1 or Thymopentin.

The beneficial effects of the present invention is: the invention discloses Thymopeptide and preparing reversing tumor associated gene mutation Reagent in application, after patient is intervened with Thymopeptide can the in early days sudden change of reversing tumor related gene, lessen the curative effect The Sudden Changing Rate of instruction mutant gene, the Sudden Changing Rate of prognosis instruction sudden change and the Sudden Changing Rate of recurrence monitoring related gene, and to tumor Patient has curative effect, alleviates patients symptomatic, extends the life cycle of patient, provides new targeting medicine for clinical treatment tumour Thing.

Accompanying drawing explanation

In order to make the purpose of the present invention, technical scheme and beneficial effect clearer, the present invention provides drawings described below:

Fig. 1 is No. 007 sample thymosin patients before and after intervention EGFR gene exons 19 deletion mutation detection figure.

Fig. 2 is No. 033 sample thymosin patients before and after intervention apc gene exons 15 point mutation detection figure.

Fig. 3 is No. 085 sample thymosin patients before and after intervention BRCA2 gene extron 11 deletion mutation detection figure.

Fig. 4 is No. 093 sample thymosin patients before and after intervention EGFR gene exon 21 abrupt climatic change figure.

Fig. 5 is No. 149 sample thymosin patients before and after intervention KRAS gene extron 2 abrupt climatic change figures.

Fig. 6 is No. 165 sample thymosin patients before and after intervention KRAS gene extron 3 abrupt climatic change figures.

Fig. 7 is No. 207 sample thymosin patients before and after intervention C-KIT exon 9 abrupt climatic change figures.

Fig. 8 is No. 243 sample thymosin patients before and after intervention BRCA15382 site mutation detection figures.

Fig. 9 is No. 297 sample thymosin patients before and after intervention PTEN exon 6 mutational site detection figures.

Figure 10 is No. 350 sample thymosin patients before and after intervention PIK3CA exon 9 mutational site detection figures.

Figure 11 is No. 411 sample thymosin patients before and after intervention NOEY26141 site detection figures.

Figure 12 is No. 600 sample thymosin patients before and after intervention EGFR exons 19 abrupt climatic change figures.

Figure 13 is No. 633 sample thymosin patients before and after intervention BRCA2 exons 11 abrupt climatic change figures.

Figure 14 is No. 749 sample thymosin patients before and after intervention EGFR exon 21 abrupt climatic change figures.

Figure 15 is No. 801 sample thymosin patients before and after intervention KRAS exon 2 abrupt climatic change figures.

Figure 16 is No. 936 sample thymosin patients before and after intervention KRAS exon 3 abrupt climatic change figures.

Figure 17 is thymosin patients before and after intervention β-tubulin abrupt climatic change figure.

Figure 18 is thymosin patients before and after intervention GAPDH abrupt climatic change figure.

Detailed description of the invention

Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.In embodiment unreceipted specifically The experimental technique of condition, generally according to normal condition, such as Molecular Cloning: A Laboratory guide (third edition, J. Pehanorm Brooker etc. writes) Described in condition, or according to the condition proposed by manufacturer.

The application in terms of reversing tumor extreme early sudden change of embodiment 1 Thymopeptide

Enter to organize without clinical symptoms personnel 1000, wherein male 500, women 500, respectively extraction 5mL venous blood, use Heparin anti-coagulating, extracts plasma DNA, and DNA extraction uses QIAGEN company test kit, and article No. is 51183, and extracting method is pressed Test kit description operates.

Proto-oncogene and the catastrophe of antioncogene in Detection and Extraction DNA, the proto-oncogene of detection has EGFR, KRAS, C-KIT and PIK3CA, the antioncogene of detection has P53, APC, BRCA1, BRCA2, PTEN and NOEY2, is correlated with non-cancer simultaneously Gene β-tubulin and GAPDH is comparison, and detection site and detection primer are as shown in table 1:

Table 1. detects the primer sequence of proto-oncogene and Tumor Suppressor Gene Mutations

Detection system is as shown in table 2:

Table 2. detection system

Reaction system forms	Add volume (μ L)
		10 × PCR Buffer is (containing MgCl₂)(Roche)	2.0

MgCl₂(Roche)	0.4
		dNTP(Roche)	0.5
Eva-green(Roche)	1.0
		10 μm ol primers (synthesis of Jie Ji Shanghai, the English Weihe River)	1.0
Template(blood plasma extracts DNA or sun control)	1.0
		Taq enzyme (Roche, HS TAQ)	0.2
H₂O	13.9

Testing conditions is as shown in table 3, and detection is at RocheCarry out on 480 instrument.

Table 3. testing conditions

Then with plasma dna as template, the sequence provided in table 1 is primer, by the reaction system in table 2 and table 3 Reaction condition to 1000 parts of pattern detection, and respectively with EGFR after testing, KRAS, C-KIT, PIK3CA, P53, APC, BRCA1, Genomic DNA unmutated for BRCA2, PTEN, NOEY2, β-tubulin and GAPDH is contrast template.Testing result shows, 1000 parts of samples there are 16 parts of pattern detection go out gene mutation, wherein male 10, women 6, the most as shown in table 4.

4.1000 parts of pattern detection results of table

Sample number	Mutational site	Follow-up clinical detects	Diagnosis
				007	EGFR exons 19	PET-CT	Adenocarcinoma of lung
033	Apc gene exons 15	PET-CT	Adenomatous polyp
				085	BRCA2 exons 11	Molybdenum target	Cystic hyperplasia
093	EGFR exon 21	PET-CT	Adenocarcinoma of lung
				149	KRAS exon 2	PET-CT	Adenomatous polyp
165	KRAS exon 3	PET-CT	Adenomatous polyp
				207	C-KIT exon 9	Gastroscope	Stomach severe atypical hyperplasia

243	BRCA15382 site	Molybdenum target	Breast nodule
				297	PTEN exon 6	PET-CT	Adenomatous polyp
350	PIK3CA exon 9	Gastroscope	Adenocarcinoma of stomach
				411	NOEY26141 site	Gastroscope	Adenocarcinoma of stomach
600	EGFR exons 19	PET-CT	Adenocarcinoma of lung
				633	BRCA2 exons 11	Molybdenum target	Cystic hyperplasia
749	EGFR exon 21	PET-CT	Adenocarcinoma of lung
				801	KRAS exon 2	PET-CT	Adenocarcinoma of lung
936	KRAS exon 3	PET-CT	Adenomatous polyp

As can be seen from Table 4, the sample of numbering 007, EGFR exons 19 deletion mutation (Fig. 1 a)；The sample of numbering 033 Apc gene exons 15 point mutation (Fig. 2 a)；The sample B RCA2 exons 11 deletion mutation (Fig. 3 a) of numbering 085；Numbering 093 Number sample EGFR gene exon 21 occur L858R suddenly change (Fig. 4 a)；The sample KRAS gene extron 2 of numbering 149 Raw G12R sudden change (Fig. 5 a)；The sample KRAS gene extron 3 of numbering 165 occurs O61H to suddenly change (Fig. 6 a)；Numbering 207 6 bases of sample C-KIT exon 9Ala502-Tyr503 section repeat to suddenly change (Fig. 7 a)；The sample B RCA1 base of numbering 243 Because C(Fig. 8 a is inserted in 5382 sites)；The sample PTEN gene extron 6 of numbering 297 is undergone mutation and (is encoded sub-179-180 to lack Lose 4bp) (Fig. 9 a)；The sample PIK3CA gene extron 9E542K of numbering 350 is sported AAA(Figure 10 a by GAA)；Numbering Sample NOEY2 gene 6141G > A(Figure 11 a of No. 411)；The sample EGFR exons 1 9A755D sudden change of numbering 600, by third Histidine mutations is aspartic acid (Figure 12 a)；6174 disappearances (figure of the sample B RCA2 gene extron 11 of numbering 633 13a)；The sample EGFR gene exon 21 of numbering 749 the 2576th is sported G(Figure 14 a by T)；The sample that numbering 801 KRAS gene extron 2 undergos mutation (G12V(GGT → GTT)) (Figure 15 a)；The sample KRAS gene extron 3 of numbering 936 Undergo mutation (O61L(CAA---CTA)) (Figure 16 a).Detecting non-cancer associated gene β-tubulin and GAPDH, result shows simultaneously Show that non-cancer associated gene β-tubulin and GAPDH is not for undergo mutation (Figure 17 a and Figure 18 a).

After obtaining above-mentioned testing result, the sample of detection sudden change is made Clinical detection, detection method and detection knot further Fruit is as shown in table 4.Showing No. 007 pattern detection, in pulmonary, blade tip end has lesser tubercle 4mm, warp after further aspiration biopsy Pathology is verified as adenocarcinoma of lung, and intramuscular injection Thymopentin (Hainan Zhonghe Pharmaceutical Co., Ltd, traditional Chinese medicines quasi-word H10970237) is each 1mg, once in a week, 3 months is a course for the treatment of, detects EGFR exons 19, outside result display EGFR after terminating lasting 3 courses for the treatment of The sudden change of aobvious son 19 disappears (Fig. 1 b)；No. 033 sample PET-CT detection is found to have Semen Glycines size polyp, pathological diagnosis after aspiration biopsy For adenomatous polyp, then intervening with Thymopentin, interference method, with No. 007 sample, rechecks apc gene after half a year Exons 15, result display apc gene exons 15 sudden change disappears；Aspiration biopsy after No. 085 sample molybdenum target detection, pathological diagnosis For cystic hyperplasia, then intervening with Thymopentin, interference method, with No. 007 sample, rechecks BRCA2 exon after half a year 11, result display BRCA2 exons 11 sudden change disappears (Fig. 3 b), and hypertrophy disappears simultaneously；No. 093 sample PET-CT detection has netted Shade, is verified as adenocarcinoma of lung through pathology after further aspiration biopsy, then intervenes with Thymopentin, and interference method is with 007 Number sample, rechecks EGFR exon 21 after half a year, result display EGFR exon 21 sudden change disappears (Fig. 4 b)；No. 149 samples PET-CT detection has diameter 1cm shade, is verified as adenomatous polyp through pathology, then uses thymus after further aspiration biopsy Pentapeptide is intervened, and interference method, with No. 007 sample, rechecks KRAS exon 2 after half a year, result display KRAS exon 2 is dashed forward Become and disappear (Fig. 5 b)；No. 165 sample clinical diagnosises are adenomatous polyp, then intervene with Thymopentin, intervention side Method, with No. 007 sample, rechecks KRAS exon 3 after half a year, result display KRAS exon 3 sudden change disappears (Fig. 6 b)；No. 207 samples This clinical diagnosis is stomach severe atypical hyperplasia, intervenes with Thymopentin, and interference method is with No. 007 sample, multiple after half a year Inspection C-KIT exon 9, result display C-KIT exon 9 sudden change disappears (Fig. 7 b)；No. 243 sample clinical diagnosises are mammary gland knot Joint, intervenes with Thymopentin, and interference method, with No. 007 sample, rechecks BRCA15382 position after half a year, result shows The sudden change of BRCA15382 position disappears (Fig. 8 b)；No. 297 sample clinical diagnosises are adenomatous polyp, do with Thymopentin In advance, interference method, with No. 007 sample, rechecks PTEN exon 6, result display PTEN exon 6 sudden change disappearance (figure after half a year 9b)；No. 350 sample clinical diagnosises are adenocarcinoma of stomach, intervene with Thymopentin, and interference method is with No. 007 sample, multiple after half a year Inspection PIK3CA exon 9, result display PIK3CA exon 9 sudden change disappears (Figure 10 b)；No. 411 sample clinical diagnosises are gastric gland Cancer, intervenes with Thymopentin, and interference method, with No. 007 sample, rechecks NOEY2 gene the 6141st after half a year, result shows Show that NOEY2 gene the 6141st sudden change disappears (Figure 11 b)；No. 600 sample clinical diagnosises are adenocarcinoma of lung, do with Thymopentin In advance, interference method, with No. 007 sample, rechecks EGFR exons 19, result display EGFR exons 19 sudden change disappearance (figure after half a year 12b)；No. 633 sample clinical diagnosises are cystic hyperplasia, intervene with Thymopentin, and interference method is with No. 007 sample, half a year Rear reinspection BRCA2 exons 11, result display BRCA2 exons 11 sudden change disappears (Figure 13 b)；No. 749 sample clinical diagnosises are Adenocarcinoma of lung, intervenes with Thymopentin, and interference method, with No. 007 sample, rechecks EGFR exon 21 after half a year, result shows EGFR exon 21 sudden change disappears (Figure 14 b)；No. 801 sample clinical diagnosises are adenocarcinoma of lung, intervene with Thymopentin, intervene Method, with No. 007 sample, rechecks KRAS exon 2 after half a year, result display KRAS exon 2 sudden change disappears (Figure 15 b)；936 Number sample clinical diagnosis is adenomatous polyp, intervenes with Thymopentin, and interference method is with No. 007 sample, after half a year Rechecking KRAS exon 3, result display KRAS exon 3 sudden change disappears (Figure 16 b).Detect after the interference of above-mentioned sample Thymopentin Non-cancer associated gene β-tubulin and GAPDH, result shows that non-cancer associated gene β-tubulin and GAPDH is not for undergo mutation (Figure 17 b and Figure 18 b).

Can obtain from above-mentioned testing result, give immunomodulator thymosin at precancerous stage, can effectively reverse Tumour associated gene mutation, even can reverse histologic lesion in early days, but on non-cancer associated gene without impact.

The application in terms of reversing tumor curative effect instruction related mutation of embodiment 2 Thymopeptide

Enter to organize tumor patient 10 example, random packet, match two-by-two, wherein pulmonary carcinoma 4 example, intestinal cancer 4 example, breast carcinoma 2 example.Enter group Tumor tissues under corrective surgery rear cutout, extracts tissue DNA, and extracting method is carried out by test kit description, DNA extraction kit Purchased from QIAGEN company, article No. is 51304.Then with extract DNA as template, utilize the reaction in the primer in table 1, table 5 System and following reaction condition screen the mutational site that 10 example patients are special, and (method is quoted from open to detect its sudden change abundance Number it is the Chinese patent of CN103397102A).

Reaction condition is as follows:

First stage: enzyme activition stage, temperature is set to 95 DEG C and carries out 5min；

Second stage: PCR expands the stage, and temperature is set to 95 DEG C and carries out 10s, and temperature is set to 60 DEG C and carries out 15s, temperature It is set to 72 DEG C and carries out 25s, and circulate 50 times；

Phase III: fusion processes, temperature is set to 95 DEG C and carries out 1min, and temperature is set to 40 DEG C and carries out 1min, temperature Being set to 65 DEG C and carry out 1s, temperature is set to 95 DEG C of detection fluorescence per second 40 times；

Fourth stage: cooling procedure, temperature is set to 40 DEG C and carries out 10s.

Table 5. reaction system

Its selection result is as shown in table 6:

Table 6.10 example patient mutations's the selection result, therapeutic scheme and therapeutic effect

After screening treating 10 example patients, therapeutic scheme is as shown in table 6, usage and consumption: Thymopentin is muscle Injection (Hainan Zhonghe Pharmaceutical Co., Ltd, traditional Chinese medicines quasi-word H10970237) 1mg every time, once in a week, 3 months is a course for the treatment of, Continue 3 courses for the treatment of.Iressa (AstraZeneca), on an empty stomach or with food with clothes, 250mg(1 sheet), 1 time on the 1st, 3 months be a treatment 3 courses for the treatment of of Cheng Chixu.Erlotinib (Roche Group), 150mg/ day, the most first 1 hour or take food latter 2 hours be administered orally, 3 Within individual month, it is to continue 3 courses for the treatment of course for the treatment of.Erbitux (Merck KGaA Xue Lannuo company), instils, and initial dose is 400mg/m², 120 minutes instillation time, drip speed and should control within 5mL/min.Maintenance dose is one week 250mg/m², the instillation time is no less than 60 minutes, once in a week, 3 months be a course for the treatment of, continues 3 courses for the treatment of.5-fluorouracil (An Kerui), 500～600mg, often Week 1 time, 3 months is a course for the treatment of, continues 3 courses for the treatment of.Trastuzumab (Roche Group), vein inputs, and initial loading dose is 4mg/ Kg, angular vein input in 90 minutes, consumption is 2mg/kg weekly, once in a week, within 3 months, is a course for the treatment of, continues 3 courses for the treatment of；Peace Consoling agent is normal saline.Detect the abundance of corresponding gene sudden change after treatment again, and add up the situation of change of Sudden Changing Rate, result such as table Shown in 6.As shown in Table 6, No. 1 patient's EGFR exon 21 point mutation declines 50%, and aobvious outside compare therewith No. 5 patient EGFR Son 21 point mutation only decline 20%；No. 3 patient's EGFR exons 19 deletion mutation declines 60%, and compare therewith No. 7 patients EGFR exons 19 lacks decline 30%；No. 2 patient's TP53 exon 5 point mutation declines 45%, and compare therewith No. 10 patients TP53 exon 5 point mutation only declines 18%；No. 4 patient's PTEN exon 6 deletion mutation declines 53%, and compare therewith No. 6 The sudden change decline 50% of patient's PTEN exon 6；No. 8 patient's BRCA2 exons 11 deletion mutation declines 80%, and compare therewith 9 Number patient's BRCA2 exons 11 deletion mutation only declines 50%.And visiting experiment patient 5 years, result shows, thymosin is done The survival of patients phase of prognosis does not carries out the patient of thymosin intervention.

The above results shows, thymosin intervenes the sudden change of the relevant canceration gene of reversible tumor efficiency instruction, by reversing Relevant mutational site has certain indicative function to curative effect.

The application in terms of reversing tumor prognosis instruction related mutation of embodiment 3 Thymopeptide

Enter to organize TNM by stages, M primary breast cancer patient 6, all there is lymphatic metastasis in various degree, all have BRCA2, NOEY2 Sudden change and HER2 gene amplification.It is randomly divided into 2 groups, often organizes 3 people.Injecting normal saline (comfort while one group of sequential therapy Agent), another group sequential therapy while inject Thymopentin (purchased from Hainan Zhonghe Pharmaceutical Co., Ltd, the quasi-word of traditional Chinese medicines H10970237), each 1mg, once in a week, 3 months is a course for the treatment of, continues 3 courses for the treatment of, after the course for the treatment of terminates, follows the tracks of and follows up a case by regular visits to 1 Year.And extract patient's 2mL whole blood anticoagulant heparin the most respectively, and extract plasma DNA, DNA extraction is normally Bright book is carried out, and test kit is purchased from QIAGEN company, and article No. is 51183, then the primer in table 1, the reaction system in table 5 and reality Execute BRCA2 and NOEY2 Sudden Changing Rate situation of change in the reaction condition detection blood plasma in example 2.Result shows, injection Thymopentin Intervention group Sudden Changing Rate averagely declines 38%, and matched group Sudden Changing Rate averagely declines 5%, intervention group survival rate 100%, and matched group life Deposit rate 33.3%.Result shows, thymosin can reverse metastasis cancer cell and be correlated with the mutation rate of canceration gene, and it is raw to improve patient Deposit rate.

The application in terms of reversing tumor recurrence monitoring related mutation of embodiment 4 Thymopeptide

Enter after group reaches to treat terminal and detect the most N/R patients with gastric cancer 60 without any clinical symptoms, iconography in 1 year, Post operation extracts paraffin organization DNA, and DNA extraction by specification is carried out, and test kit is purchased from QIAGEN company, and article No. is 51183.So The rear Chinese patent utilizing Publication No. CN103397102A filters out and suddenlys change patient 2 containing TP53, and PIK3CA suddenlys change patient 2 Name, CDH1 suddenlys change patient 2, then takes patient whole blood's anticoagulant heparin, the primer in table 1, the reaction system in table 2 and embodiment Testing conditions in 3 detects each patient's corresponding gene Sudden Changing Rate.After detection, a patient in mutation type of the same race is injected thymus Pentapeptide (purchased from Hainan Zhonghe Pharmaceutical Co., Ltd, traditional Chinese medicines quasi-word H10970237), each 1mg, once in a week, within 3 months, it is one The course for the treatment of, another one patient injects the normal saline of equivalent as placebo.After 3 months, identical method is utilized to detect each The sudden change changes of contents situation of patient's correspondence mutant gene.It was found that test group Sudden Changing Rate declines is above matched group.And The matched group patient of TP53 sudden change patient confirms to recur through iconography.

Finally illustrate, preferred embodiment above only in order to technical scheme to be described and unrestricted, although logical Cross above preferred embodiment the present invention to be described in detail, it is to be understood by those skilled in the art that can be In form and it is made various change, without departing from claims of the present invention limited range in details.

Claims

1. Thymopeptide application in the reagent preparing reversing tumor patient tumors associated gene mutation, described thymosin Preparation is Thymopentin；It is described tumor-related that because proto-oncogene or antioncogene, described proto-oncogene is EGFR, KRAS, C-KIT or PIK3CA；Described antioncogene is BRCA1, BRCA2, PTEN or NOEY2；Specifically sport EGFR exons 19 A755D suddenlys change；EGFR exon 21 occurs L858R to suddenly change；There is G12R sudden change or G12V sudden change in KRAS gene extron 2； There is O61H sudden change or O61L sudden change in KRAS gene extron 3；6 bases of C-KIT exon 9 Ala502-Tyr503 section Repeat sudden change；PIK3CA gene extron 9 E542K suddenlys change；C is inserted in BRCA1 gene 5382 site；The of BRCA2 exons 11 6174 disappearances；PTEN gene extron 6 codon 179-180 lacks 4bp；NOEY2 gene 6141 G > A suddenlys change.