CN106166291A - Spleen polypeptide improves KLRK1 or LCK and treats immunosuppressant medical usage - Google Patents
Spleen polypeptide improves KLRK1 or LCK and treats immunosuppressant medical usage Download PDFInfo
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Abstract
The present invention relates to spleen polypeptide and improve KLRK1 or LCK treatment immunosuppressant medical usage, particularly spleen polypeptide application in the medicine preparing the disease that KLRK1 or LCK albumen lowly causes, the disease that described KLRK1 or LCK albumen lowly causes includes: tumor, cause pathogeny imcrobe infection, flu, asthma, bronchitis, rheumatoid arthritis, present invention additionally comprises spleen polypeptide and with cyclophosphamide combined medication application in preparing antitumor drug.
Description
Technical field
The present invention relates to spleen polypeptide and improve killing cell agglutinin sample receptor K1 (KLRK1) or lymphocyte specific protein
Tyrosine kinase (LCK) treatment immunosuppressant medical usage.It is expected to reduce caused disease for various because KLRK1 or LCK expresses
Sick treatment.
Background technology
1, act in advance without antigen just can direct killing tumor and disease for natural killer cell (natural killer, NK)
The target cell that poison infects, because possessing this functional characteristics, NK cell monitors and during early stage anti-infectious immunity at immunity of organism
Play an important role.The distribution of NK cell surface kills activated receptor and killerinhibitoryreceptor, and the interaction by two kinds of receptors is true
Protect NK cell and can kill cell and the mutated tumor cell of virus infection, host self normal cell can be protected again to exempt from brokenly
Bad.NK cell surface killerinhibitoryreceptor combines with target cells histocompatibility complex class Ⅰmolecule (mhc class i molecule),
Killing suppression signal can be produced, the transmission killing signal can be blocked.For host autologous tissue cell, because of mhc class i developed by molecule
The inhibitory action normally making killerinhibitoryreceptor mediate is occupied an leading position, and shows as NK cell inactivation；For virus infect thin
Born of the same parents and tumor cell, reduce due to mhc class i developed by molecule or lack and affect the combination of killerinhibitoryreceptor and respective ligand,
So that the effect killing activated receptor is occupied an leading position, show as NK cell activation and produce lethal effect.Recent study
Showing, human cell surface's Inhibitory receptor can be divided into three families:
First family is killer cell immunoglobulin-like receptors (Killer Ig-like Receptors KIR),
Have a type membrane protein receptor of two to three immunoglobulin like domain in structure for extracellular fragment, part is that human leucocyte resists
Former class Ⅰmolecule (HLA class Ⅰmolecule).
Second family is immunoglobulin-like transcription product (ILT), including ILT-4, ILT-2 etc..
3rd family is for killing cell agglutinin sample receptor (Killer cell Lectin-like
ReceptorsKLR), all members are positioned the NK complex (NKC) of No. 12 chromosomes, are subject to including killing cell agglutinin sample
Body K1 (KLRK1), CD94/NKG2, NKR-P1A, KLRF1, CD69, CLEC-2 etc..
The construction features of inhibitive ability of immunity receptor is that intracellular section contains relevant the pressing down of one or more immunity receptor tyrosine
Property motif processed (immunoreceptor tyrosine-based inhibitory motifs ITIM), ITIM is by 6 ammonia
The conserved sequence (I/L/VXYXXL/V, X represent arbitrary aminoacid) of base acid composition.When inhibitive ability of immunity receptor is even with respective ligand
During connection, there is tyrosine phosphorylation in ITIM, then with SHP1 Protein-tyrosine-phosphatase or/and the knot of SPIP inositol monophosphate enzyme
Close, produce the negative regulator effect of cell activation.
Owing to inhibitive ability of immunity receptor plays a significant role in cytoactive regulates, especially this class of discovered in recent years is divided
Son all has expression at cytotoxic T lymphocyte and dendritic cell (Dendritic cells, DC) surface so that it is have important
Theory study meaning, be the heat subject in current immunology, cell and the field such as molecular biology and oncology.
2, Lymphocyte-specific protein-tyrosine kinase (Lymphocyte specific protein tyrosine
Kinase, Lck) it is the protein tyrosine kinase of Src family member, it is primarily present in T lymphocyte, participates in sending out of T cell
The signal transduction process educate, break up, activated.The display of many research both at home and abroad: the unconventionality expression of Lck and regulation and the canceration of cell
Closely related, as human lymphocyte and non-lymphocyte malignant tumor all have Lck unconventionality expression or activation, at transgenic mice,
The expression of Lck causes thymus neoplasms.In Lck domain catalytic domain contain two be available for regulation tyrosine residues (tyr394 and
Tyr505), tyr394 is an autophosphorylation site, Lck kinase activity can be made to raise by self phosphorylation；tyr505
Phosphorylation then can make Lck kinase activity down-regulation, if with after phenylalanine displacement tyrosine (Y505F) that can not be phosphorylated,
The lasting rising of Lck kinase activity then occurs；Lck can promote STAT5b tyrosine phosphorylation, thus activates STAT5b, promotes
STAT5b Yu DNA adhesion increases.More than research shows: LCK plays a significant role in tumor and disease of immune system.
3, spleen peptide injection is a kind of polypeptide drugs that are that Jilin Fengsheng Pharmaceutical Co., Ltd. produces and that listed.Its
Polypeptide is that the molecular weight that calf spleen extract is made is less than 6,000 daltonian polypeptide, free amino acid, nucleic acid, total sugar
Aseptic aqueous solution.This product every 1mL 4.0mg Han polypeptide, is 5.0mg containing free amino acid, is 1.0mg containing nucleic acid, the lowest containing total sugar
In 100 μ g.Spleen peptide injection has dual regulation to human body immune function, it is possible to rectifier body immunity function is disorderly, tool
Have and activate and the effect of enhancing body non-specific immunity, it is possible to promote T lymphocyte maturation and non-sensitization lymph can be made
Cell-stimulating becomes primed lymphocyte, thus improves lymphocyte immunologic function, triggers and infection is supported by enhancing body
Drag.Spleen peptide injection can also the generation of inducing interferon, the directly synthesis of prevention virus protein and duplication, and energy
Strengthen cell surface antigen to express, promote the cytotoxic activity of NK cell, regulate lymphocyte and macrophage function, can be obvious
Improve Immune Function.Spleen peptide injection can stimulate proliferation of bone marrow cells, produces a large amount of leukocyte, makes hemopoietic function
Carried.Additionally, spleen peptide injection can suppress cell glycolysis with non-toxic, make to be characterized with height glycolysis
Tumor cell lacks energy source, causes tumor cell metabolic process generation obstacle, and prevention G0, G1 phase tumor cell can not be to increasing
Grow, division stage, develops, thus reaches anticancer effect.
Spleen peptide injection can be clinically used for constitutional and secondary cell immunodeficiency (as eczema, platelet subtract
Less, many subinfections syndrome etc.), respiratory tract and pulmonary infection, the leukopenia that can cause at treatment chemicotherapy, white blood
Disease, reproducibility obstacle anemia, lymphoma and other malignant tumor, improve tumor patient cancerate matter, improve postoperative or patient with severe symptoms
During physical weakness, auxiliary uses.
The present invention uses immunosuppressant and lotus tumor scale-model investigation its concrete improvement immunity and and chemotherapy drugs in combination
During the mechanism of action of medication, it has unexpectedly been found that spleen polypeptide can kill cell agglutinin sample receptor K1 by improving
(KLRK1) or the expression of lymphocyte specific protein tyrosine kinase (LCK), reach to treat immunosuppressant and antineoplastic is made
With, thus be expected for the various treatments expressing reduction associated diseases because of KLRK1 or LCK.
Summary of the invention
The invention provides spleen polypeptide and can expect for treating and preventing various because of KLRK1 or the LCK caused disease of expression reduction
Sick medical usage.
It is a discovery of the invention that spleen polypeptideCan effectively improve caused by cyclophosphamide because ofThe expression of KLRK1, LCK albumen
Decline the immunosuppressant caused.Therefore spleen polypeptide may be used for treatmentCauseThe expression decline of KLRK1, LCK albumen is caused
Immune disease and tumor-related illness.
To this end, the present invention provides the new medicinal usage of spleen polypeptide.
The new medicinal usage of spleen polypeptide of the present invention includes:
The application in the medicine preparing the disease that KLRK1 or LCK albumen lowly causes of the spleen polypeptide.
Described application, including because of the treatment of the low associated diseases of KLRK1 or LCK.Caused by KLRK1 or LCK reduction abnormality
Disease includes: tumor, cause pathogeny imcrobe infection, flu, asthma, bronchitis, rheumatoid arthritis etc..
Present invention additionally comprises, spleen polypeptide and with cyclophosphamide combined medication application in preparing antitumor drug.
Present invention additionally comprises, spleen polypeptide and with cyclophosphamide combined medication preparation improve tumour patient immunity medicine
In application.
Spleen polypeptide of the present invention and with cyclophosphamide combined medication, it is characterised in that both proportionings are:
Spleen polypeptide 50-100 weight portion, cyclophosphamide 20-80 weight portion.
Preferably include,
Spleen polypeptide 50 weight portion, cyclophosphamide 20 weight portion.
Spleen polypeptide 50 weight portion, cyclophosphamide 40 weight portion.
Spleen polypeptide 50 weight portion, cyclophosphamide 80 weight portion.
Spleen polypeptide 100 weight portion, cyclophosphamide 20 weight portion.
Spleen polypeptide 100 weight portion, cyclophosphamide 40 weight portion.
Spleen polypeptide 100 weight portion, cyclophosphamide 80 weight portion.
To this end, the present invention further provides the compound medicament composition of spleen polypeptide and cyclophosphamide, in described compositions two
Person's proportioning is: spleen polypeptide 50-100 weight portion, cyclophosphamide 20-80 weight portion.
The compound medicament composition of the present invention, it may be necessary to add pharmaceutically acceptable carrier.
In above proportioning, when producing can by kilogram in units of, or in units of ton, small-scale production can also with gram
Or milligram is unit, weight can increase or reduce, but the constant rate of the raw medicinal herbs weight proportion between each composition.
The ratio of above proportioning obtains through science screening, for special circumstances, such as serious symptom or mild, fat or thin
Little, the proportioning of the amount of composition can be adjusted accordingly, be increased or decreased less than 100%, drug effect is constant.
The compound of the present invention, when being prepared as pharmaceutical preparation, can add pharmaceutically acceptable carrier if desired, press
Prepared by the routine techniques according to galenic pharmacy.The pharmaceutical dosage forms of the present invention, exists in a unit, described unit dose shape
Formula refers to the unit of preparation, such as every of tablet, and every seed lac capsule of capsule, granule every bag etc..These dosage forms include: tablet,
Capsule, granule, pill, powder, unguentum, sublimed preparation, powder, solution, injection, suppository, spray, drop, patch.This
The preparation of invention, preferably peroral dosage form, such as: capsule, tablet, oral liquid, granule, pill, powder, sublimed preparation, unguentum
Deng.
The compound of the present invention, the preparation of its oral administration can be containing conventional excipient, such as binding agent, filling
Agent, diluent, tablet agent, lubricant, disintegrating agent, coloring agent, flavoring agent and wetting agent.
The form of oral liquid can be such as aqueous or oily suspensions, solution, Emulsion, syrup or elixir,
Or can be the compounding dry products of a kind of available water before use or other suitable carrier.This liquid preparation can contain
Conventional additive, such as suspending agent, such as sorbitol, syrup, methylcellulose, gelatin, hydroxyethyl cellulose, carboxymethyl fibre
Dimension element, aluminium stearate gel or hydrogenated edible fats, emulsifying agent, such as lecithin, anhydro sorbitol monooleate or Arab
Glue；Non-aqueous carrier (they can include edible oil), the oily ester of the such as ester of almond oil, fractionated coconut oil, such as glycerol,
Propylene glycol or ethanol；Preservative, such as para hydroxybenzene methyl ester or propyl p-hydroxybenzoate or sorbic acid, and if it is required,
Can be containing conventional flavouring agent or coloring agent.
For injection, the fluid unit dosage form of preparation contains active substance and the sterile carrier of the present invention.According to carrier
And concentration, this compound can be suspended or dissolve.The preparation of solution is typically by active substance is dissolved in a kind of load
In body, filter-sterilized before being loaded into a kind of suitable bottle or ampoule, then seal.Such as a kind of local anesthesia of adjuvant
Agent, preservative and buffer agent can also be dissolved in this carrier.In order to improve its stability, can be by this after loading bottle
Plant compositions frost, and under vacuo water is removed.
Beneficial effects of the present invention is further illustrated below by way of experimental data:
During research spleen polypeptide with the cyclophosphamide combined medication effect to lotus tumor model, it has been found that:
1, spleen polypeptide and the cyclophosphamide combined medication effect to tumor-bearing mice survival rate
As it is shown in figure 1, the mortality rate of mice is had no significant effect by CY20mg/kg each dosage group substantially, only CY20mg/kg agent
There is 17% mortality rate at the 16th day in amount group mice.CY40mg/kg and death occurs with SP synergy group when the 11st day
Phenomenon, wherein the change of CY40mg/kg independent medication group mortality rate is the most obvious, and all animals are substantially the most dead in 21 days；When
CY40mg/kg respectively with SP drug combination after, the survival rate of mice has extended, during the 12nd day to the 21st day, drug combination
The survival rate of group is significantly raised compared with CY40mg/kg independent medication group.CY80mg/kg and with SP drug combination after, due to CY show
The toxic action write causes the survival rate of all group mices to be below 21 days, but the life span of CY80+SP100 dosage group
Be considerably longer than the independent medication of CY80mg/kg in 21 days 14 days.This experimental result explanation CY80mg/kg with CY40mg/kg individually makees
Strong by group toxicity, dead mouse phenomenon is obvious, when with SP drug combination after, life span and survival rate all make moderate progress, its
In the most notable with SP100mg/kg dosage combinations medication improvement result.
2, spleen polypeptide (SP) and the effect to tumor-bearing mice body weight of cyclophosphamide (CY) drug combination
The effect (n=10-12) to tumor-bearing mice body weight of table 1 drug combination
*, compare with blank group, P < 0.05；#, compares with model group, P < 0.05；
As shown in table 1, after giving CY treatment, the body weight of mice is all remarkably decreased.When CY dosage is 20mg/kg dosage,
Individually medication group Mouse Weight relatively naive mice significantly reduces, after SP drug combination, and the body weight of mice and blank group and mould
Type group is all without significant difference.As in figure 2 it is shown, after CY20 Yu SP drug combination, the body weight of mice starts appearance after being administered on the 6th day
Rise trend, final and normal mouse body weight no significant difference.Illustrate that CY20mg/kg is controlled by CY20mg/kg with SP drug combination
The Mouse Weight produced during treatment reduces phenomenon an improvement result, and SP50mg/kg dosage group and SP100mg/kg dosage group
Action effect is close.
As shown in table 1, after giving CY40mg/kg treatment, the body weight of mice is all remarkably decreased and relatively CY20mg/kg dosage
Group reduces substantially.CY40mg/kg dosage respectively with SP drug combination after, the body weight of mice is had certain improvement, wherein
SP100mg/kg dosage group Mouse Weight there was no significant difference with model group Mouse Weight.As in figure 2 it is shown, CY40mg/kg dosage
The body weight change trend of group and CY40mg/kg with SP50mg/kg drug combination group mice is basically identical, but CY40mg/kg
After treating 9 days, certain recovery trend is had with the body weight of SP100mg/kg drug combination group mice.CY40mg/kg dosage is described
Mouse Weight produces significantly impact, and the body weight to mice that adds of SP100mg/kg has some improvement.
As shown in table 1, similar with CY40mg/kg dosage group, after giving CY80mg/kg treatment, the body weight of mice is the most aobvious
Write and decline.After drug combination, the body weight of mice there is is certain improvement, but improves DeGrain.As in figure 2 it is shown, CY80mg/
The body weight change trend of kg dosage group and CY80mg/kg with SP drug combination group mice is basically identical, and the addition base of SP is described
The toxic action in basis caused CY80mg/kg is not improved effect.
In sum, CY long-term prescription is notable to mouse toxicity effect, Mouse Weight change substantially, when with SP drug combination
Body weight to mice has certain improvement afterwards, and wherein Mouse Weight is had some improvement by SP100mg/kg drug combination.
3, the drug combination effect to tumor-bearing mice organ coefficient
The effect (n=10-12) to tumor-bearing mice organ coefficient of table 2 drug combination
*, compare with blank group, P < 0.05；#, compares with model group, P < 0.05；
Shown in table 2, through labor, find that between each group of experiment, liver and renal index are substantially without the difference of significance
Different, illustrate that cyclophosphamide and the medication alone or in combination of spleen polypeptide all do not produce obvious liver and Toxicity of Kidney.Study simultaneously
After finding model group mouse inoculation tumor, more blank group of spleen index has a certain degree of increase.The analysis found that, blank group is little
The index and spleen index of Mus is big compared with the index that document is reported, this is likely due in therapeutic process tail vein injection saline continuously
And then induce strong inflammatory reaction and produce[22,23].The addition of cyclophosphamide is notable on thymus index impact, various dose
After spleen polypeptide and cyclophosphamide combined medication, thymus index is had improvement to a certain extent.Therefore we will be discussed in detail difference
Dose medication regimen is to mouse spleen and the effect of thymus index.
In table 2, CY20mg/kg dosage group is close with CY20mg/kg and SP50mg/kg drug combination group spleen index, and this is described
The effect on spleen index that adds of the lower SP50mg/kg of dosage CY effect is substantially without impact.But the addition of SP100mg/kg then shows
Write reduces spleen index, alleviates spleen swelling phenomenon.We have found that this dosage independent role group and synergy group simultaneously
Thymus index and blank group and model group there are no significant difference.The cyclophosphamide thymus index to mice of this dosage is described
Substantially without impact, obvious immunosuppressive action may not produced.
Such as table 2, the index and spleen index of CY40mg/kg dosage group is closer to blank group and model group index and spleen index, but works as
After this dose Cyclophosphamide and spleen polypeptide drug combination, index and spleen index significantly lowers.Illustrate that the addition of spleen polypeptide may certain journey
Alleviate inflammatory effect on degree, and then reduce the swelling phenomenon of spleen.This dosage each effect group the most significantly reduces mice
Thymus index, illustrate that the cyclophosphamide of this dosage shows certain immunosuppressive action.But the addition of spleen polypeptide is not
Thymus index is produced the impact of significance.
Table 2 shows, the independent medication of CY80mg/kg dosage group and equal with the index and spleen index of spleen polypeptide drug combination group mice
Being in higher level, the addition of spleen polypeptide fails to alleviate the swelling phenomenon of spleen.Meanwhile each group of this dosage is likely to be due to produce
Raw significant immunosuppressant phenomenon, thymus index all significantly reduces.
4, the drug combination effect to tumor-bearing mice leukocyte count
Leukocyte count variation tendency shows, model group mice is due to the invasion of exogenous tumor cell, and then induction is white thin
Born of the same parents' number increases phenomenon；The independent long-term prescription of CY produces certain immunosuppressant phenomenon, and along with the increase of CY dosage, immunosuppressant is existing
As being consequently increased (Fig. 3).But when CY 80mg/kg individually or combines long-term prescription with SP, the leukocyte digital display in blood
Write and increase, it may be possible to the leukemia due to high dose Induced by Cyclophosphamide.The independent medication of CY20mg/kg Yu CY40mg/kg simultaneously
Leukocyte count significantly reduces, but is as the addition of spleen polypeptide, and leukocyte count all has rising to a certain extent, and spleen polypeptide is described
The immunosuppressive action that cyclophosphamide produces may there be is certain mitigation.
5, drug combination is to tumor-bearing mice gross tumor volume and the effect of weight
The effect (n=10-12) to tumor-bearing mice tumor weight of table 3 drug combination
#P < 0.05, compares with blank group.
Along with the increase of CY dosage, the effect of its suppression tumor growth strengthens the most therewith, as shown in table 3, and CY80mg/kg group
And this dosage is up to more than 90% with SP drug combination group tumor control rate, substantially completely inhibits the growth of tumor and kill
Dead tumor cell.CY20mg/kg, CY40mg/kg group and also show good suppression tumor with SP drug combination group simultaneously
Effect (suppression ratio is more than 50%).The change of gross tumor volume in our labor therapeutic process, from Fig. 4 it is found that
After starting to treat the 5th day, CY20mg/kg and relevant drug combination group, gross tumor volume variation tendency is slow, but CY40mg/
Kg to CY80mg/kg dosage and relevant drug combination group, gross tumor volume is in being decreased obviously trend.Simultaneously according to above-mentioned data,
It appeared that we are along with the increase of CY dosage, and tumor inhibitory effect is strengthened therewith, but SP with CY drug combination does not improve
The tumor inhibition effect of CY.
6, drug combination is on the impact of T cell subset proportions in tumor-bearing mice blood
Table 4 drug combination is on the impact of T cell subset proportions in tumor-bearing mice blood
* compare with blank group, P < 0.05；# compares with model group, P < 0.05；
Table 4 shows the CD3 increased in blood along with cyclophosphamide dosage+Cell dosage dependency increases.In leukocyte
20-40% is lymphocyte, and CD3+Can be used for characterizing the quantity of mature T lymphocyte in peripheral blood.Therefore explanation may be by
Cyclophosphamide long-term prescription in high dose induces leukemia to a certain extent, and CD3+ cell the most therein increases the most therewith
Add.
CD4+For helper T lymphocyte, participate in cellular immunization and the formation of delayed hypersensitivity inflammation, in cause of disease body-sensing
Dye plays an important role；And B cell proliferation can be stimulated and produce antibody, closely related with humoral immunization.Cyclophosphamide is individually used
Medicine group significance reduce CD4 in blood+Level, only CY20mg/kg with SP100mg/kg drug combination effect group necessarily shows
The CD4 that improve in blood write+Level.
CD8+Being a kind of cytotoxic T cell, it mainly directly kills target by secretion perforin, cytokine TNF etc.
Cell.Understand after data analysis in table 4, CY80mg/kg and this dosage combinations effect group, the CD8 in mouse blood+Level
Notable attenuating, the addition of spleen polypeptide does not significantly improve this effect, therefore illustrates that the cyclophosphamide of this dosage may produce notable
Immunosuppressive action, thus reduce the cellular immune level in blood.
CD4+/CD8+The change of ratio is closely related with body immunity level.Through the finishing analysis of mass data, send out
The addition of existing cyclophosphamide significantly reduces the CD4 in blood+/CD8+Ratio, illustrates that cyclophosphamide long-term prescription will be significantly
Reduce the immunity level of body.When SP100mg/kg respectively with tri-various dose drug combinations of CY after, to CD4+/CD8+Ratio
The rate effect of being improved to some extent.Illustrate that the immunity level to body that adds of spleen polypeptide has some improvement.
7, the drug combination impact on tumor-bearing mice tumor transcriptional group
Illumina 2000 is utilized to analyze, (RNAseq) analysis test method, test in the tumor tissues of each treated animal
Total mRNA, it has been carried out order-checking and content analysis.Experimentation is as follows:
Sample packet situation:
Model group；
CY40: cyclophosphamide 40mg/kg independent medication group；
CY80: cyclophosphamide 80mg/kg independent medication group；
CY40+SP100: cyclophosphamide 40mg/kg with spleen polypeptide 100mg/kg drug combination group；
CY80+SP100: cyclophosphamide 80mg/kg with spleen polypeptide 100mg/kg drug combination group；
Experimental technique:
RNAseq initial data is carried out screening strength, is analyzed comparing to each group of data between any two respectively, sieve
Select wherein diversity bigger carry out selective analysis.Then gene bigger for diversity is carried out GO annotation, diligent according to making
Can classify, filter out wherein relevant to growth of cancers suppression, inflammatory reaction, immunomodulating etc. gene.Finally according to literary composition
Offer mining analysis, filter out key protein.
Interpretation:
Table 5 each albumen expression situation in different groups
Data in upper table are in RNAseq data analysis and produce notable change (rise or decline) between each group,
And the albumen relevant to key words such as tumor, inflammation, immunomodulating.As can be seen from the table, after CY80 monotherapy,
The expression of KLRK1, LCK albumen is all decreased obviously, and illustrates to produce immunosuppressive action, induces above-mentioned and Ia albumen
The decline of expression.Meanwhile, we to the albumen variation tendency of CY80+SP100 drug combination group with individually dosed group
CY80 compares analysis, illustrates, when cyclophosphamide (CY) and spleen peptide injection drug combination, producing antitumous effect
Meanwhile, spleen peptide injection improves immunity of organisms, thus promotes antitumous effect, reduces chemotherapeutics toxic action.
By result above it was unexpectedly observed that spleen polypeptideCan effectively improve caused by cyclophosphamide because ofKLRK1, LCK egg
White expression declines the immunosuppressant caused.Thus, it was found that spleen polypeptide is in treatmentCauseThe expression of KLRK1, LCK albumen
Decline the immune disease and the treatment of tumor-related illness caused.
Accompanying drawing explanation
Fig. 1 spleen polypeptide (SP) and the cyclophosphamide (CY) effect (n=10-12) to tumor-bearing mice survival rate.
Fig. 2 drug combination effect (n=10-12) * to tumor-bearing mice body weight, compares with blank group, P < 0.05；#, with mould
Type group compares, P < 0.05；
Fig. 3 drug combination effect (n=10-12) .*P < 0.05 to tumor-bearing mice leukocyte count, compares with blank group；#P
< 0.05, new with mould group is compared (after data acquisition self administration of medication the 19th day)
The effect (n=10-12) to tumor-bearing mice gross tumor volume of Fig. 4 drug combination. (A) gross tumor volume；(B) tumor shape
State #P < 0.05, compare with blank group, 1: blank group, 2: model group, 3:CY20 group, 4:CY40 group, 5:CY80 group, 6:CY20+
SP50 group, 7:CY20+SP100 group, 8:CY40+SP50 group, 9:CY40+SP100 group, 10:CY80+SP50 group, 11:CY80+
SP100 group
Detailed description of the invention
Following example represent the practicality of the present invention, and the present invention is not limited.
Embodiment 1 spleen polypeptide improves caused by cyclophosphamide tumor-bearing mice immunosuppressive action
1.1 experimental animal model
Male Kunming strain mice 220, is randomly divided into 11 groups.Often 20 mices of group, wherein 10 are flowed according to above-mentioned experiment
Journey, put to death in the 19th day, and plucking eyeball, to take blood standby, wins the histoorgans such as thymus, spleen, liver, kidney simultaneously, weighs and calculate dirty
Device index；Remaining 10 started stopping from the 19th day and give CY, continue to raise and test its survival rate change curve.Above-mentioned mice is divided
For: blank group, model group, CY20, CY40, CY80, CY20+SP50, CY20+SP100, CY40+SP50, CY40+SP100,
CY80+SP50, CY80+SP100.In addition to blank group, each group was inoculated 0.2mL density in the 0th day in oxter, right side is 1 × 107's
S180 cell, in the 7th day, every day lumbar injection 0.1mL/10g medicine.Its empty group injecting normal saline, each group of CY
Give 20 respectively, 40,80mg/kg cyclophosphamide, CY+SP group gives 50 at the cyclophosphamide giving corresponding dosage simultaneously,
100mg/kg spleen peptide injection.
1.2 gross tumor volumes, tumour inhibiting rate method of testing
In experimentation, observe the size of mouse tumor volume every day, utilize length and width, the meter of slide gauge test tumor
Calculate the volume of tumor, and draw a day gross tumor volume change curve.After experiment terminates, win mouse tumor, weigh, and count
Calculate tumour inhibiting rate (In).
Gross tumor volume=0.5 × length × wide2
In=(model group tumor weight-administration group tumor weight) ÷ model group tumor weight
1.3 animal survival curve test methods
In therapeutic process, every day records the survival condition of mice, draws the survival rate change curve of mice[20]。
Survival rate=(this treated animal initial number-this treated animal quantity on hand)/this treated animal initial number.
1.4 experimental result
Experimental result is shown in Table 1-table 5 and Fig. 1-Fig. 3, and concrete conclusion is as follows:
1) according to experimental result, cyclophosphamide CY40mg/kg and CY80mg/kg dosage group dosage over the course for the treatment of are found
Dependent generation immunosuppressive action to a certain extent, significantly reduces the thymus index of mice, and the CD4 in blood+、CD8+、D4+/CD8+Level.
2) SP Yu CY drug combination can not improve the CY antitumor action to S180 tumor-bearing mice substantially.But can certain journey
Improve the immunity level of mice on degree, improve life span and the survival rate of mice.
3) analyzed by transcription group, it was found that spleen polypeptideCan effectively improve caused by cyclophosphamide because ofKLRK1、
The expression of LCK albumen declines the immunosuppressant (being shown in Table 5) caused.
Claims (10)
1. spleen polypeptide application in the medicine preparing the disease that KLRK1 or LCK albumen lowly causes.
Application the most according to claim 1, it is characterised in that the disease that KLRK1 or LCK albumen lowly causes includes: swollen
Tumor, cause pathogeny imcrobe infection, flu, asthma, bronchitis, rheumatoid arthritis.
3. spleen polypeptide and with cyclophosphamide combined medication application in preparing antitumor drug.
4. spleen polypeptide and with cyclophosphamide combined medication preparation improve tumour patient immunity medicine in application.
5. according to the application described in Claims 2 or 3 any one, it is characterised in that wherein spleen polypeptide and with cyclophosphamide join
Share medicine, both proportionings are: spleen polypeptide 50-100 weight portion, cyclophosphamide 20-80 weight portion.
6. spleen polypeptide and the compound medicament composition of cyclophosphamide, it is characterised in that in described compositions, both proportionings are: spleen is many
Peptide 50-100 weight portion, cyclophosphamide 20-80 weight portion.
Compositions the most according to claim 6, it is characterised in that in described compositions, both proportionings are: spleen polypeptide 50 weight
Amount part, cyclophosphamide 20 weight portion, or spleen polypeptide 50 weight portion, cyclophosphamide 40 weight portion, or spleen polypeptide 50 weight portion, ring phosphorus
Amide 80 weight portion.
Compositions the most according to claim 6, it is characterised in that in described compositions, both proportionings are: spleen polypeptide 100 weight
Amount part, cyclophosphamide 20 weight portion, or spleen polypeptide 100 weight portion, cyclophosphamide 40 weight portion, or spleen polypeptide 100 weight portion, ring
Phosphamide 80 weight portion.
Compositions the most according to claim 6, it is characterised in that described pharmaceutical composition can add medicine if desired can
The carrier accepted.
Compositions the most according to claim 6, it is characterised in that the dosage form of described compositions includes: tablet, capsule,
Granule, pill, powder, injection, suppository, spray, drop pill.
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| CN201610638104.4A CN106166291B (en) | 2016-08-05 | 2016-08-05 | Medical application of spleen polypeptide in improving KLRK1 or LCK treatment immunosuppression |
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| CN201610638104.4A CN106166291B (en) | 2016-08-05 | 2016-08-05 | Medical application of spleen polypeptide in improving KLRK1 or LCK treatment immunosuppression |
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| CN106166291B CN106166291B (en) | 2020-06-30 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2766340C1 (en) * | 2021-02-10 | 2022-03-15 | Игорь Закванович Зайцев | Peptide tolerogenic compound |
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| CN1634987A (en) * | 2004-03-10 | 2005-07-06 | 菲尔斯·杜克制药（通化）有限公司 | Spleen polypeptide extract, its preparing process and use |
| CN103006622A (en) * | 2011-09-21 | 2013-04-03 | 成都中医药大学 | New borneol use and lung cancer treatment drug composition |
| CN104127427A (en) * | 2014-07-28 | 2014-11-05 | 李健 | Anticancer synergistic composition |
| CN104147031A (en) * | 2014-07-28 | 2014-11-19 | 李健 | Antitumor pharmaceutical composition containing aesculin |
| CN105342996A (en) * | 2015-11-24 | 2016-02-24 | 深圳市艾民医药科技有限公司 | Preparation method for spleen polypeptide injection |
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2016
- 2016-08-05 CN CN201610638104.4A patent/CN106166291B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1634987A (en) * | 2004-03-10 | 2005-07-06 | 菲尔斯·杜克制药（通化）有限公司 | Spleen polypeptide extract, its preparing process and use |
| CN103006622A (en) * | 2011-09-21 | 2013-04-03 | 成都中医药大学 | New borneol use and lung cancer treatment drug composition |
| CN104127427A (en) * | 2014-07-28 | 2014-11-05 | 李健 | Anticancer synergistic composition |
| CN104147031A (en) * | 2014-07-28 | 2014-11-19 | 李健 | Antitumor pharmaceutical composition containing aesculin |
| CN105342996A (en) * | 2015-11-24 | 2016-02-24 | 深圳市艾民医药科技有限公司 | Preparation method for spleen polypeptide injection |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2766340C1 (en) * | 2021-02-10 | 2022-03-15 | Игорь Закванович Зайцев | Peptide tolerogenic compound |
| WO2022173328A1 (en) * | 2021-02-10 | 2022-08-18 | Игорь Закванович Зайцев | Tolerogenic peptide compound |
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| Publication number | Publication date |
|---|---|
| CN106166291B (en) | 2020-06-30 |
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Address after: 135000 no.4455 Jianguo Road, Meihekou City, Tonghua City, Jilin Province Applicant after: Rongzhi Fengsheng Pharmaceutical Co.,Ltd. Applicant after: SHENYANG PHARMACEUTICAL University Address before: 135000 Jianguo Road, Jilin, Meihekou, No. 4455 Applicant before: JILIN FENGSHENG PHARMACEUTICAL Co.,Ltd. Applicant before: SHENYANG PHARMACEUTICAL University |
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