CN110115767A - The method of immunomodulator joint inspection point inhibitor for treating cancer - Google Patents

Abstract

The present invention provides a kind of use in conjunction 1) immunomodulator, 2) checkpoint inhibitor and 3) discharge cancer antigen therapy for treatment of cancer patient method.In some respects, the immunomodulator can be anti-CD3 immunotoxin, and the checkpoint inhibitor inhibits the interaction of PD-1 and PD-L1, and the cancer antigen release therapy is radiotherapy.

Description

The method of immunomodulator joint inspection point inhibitor for treating cancer

Background of invention

Invention field

The invention mainly relates to use in conjunction 1) immunomodulator, 2) checkpoint inhibitor and the 3) treatment of release cancer antigen The method of method (such as radiotherapy) treating cancer patient.Specifically, the immunomodulator can be anti-CD3 immunotoxin, the inspection Make an inventory of the interaction that inhibitor inhibits PD-1 and PD-L1.

Background of invention

Interaction between PD-1 albumen on the PD-L1 albumen and T cell of cell surface reduces the function of T cell And prevent immune system attack cell.Although this interaction is very useful to protection normal cell, some cancer cells It can be in its surface expression PD-L1 and the normal cell that disguises oneself as is not by immune system attack.This cancer cell energy for being loaded with PD-L1 It is enough to escape immune clearance whereby.To solve this problem, developed " checkpoint inhibitor " (majority be immune antiboidy) can be with The interaction of PD-L1 and PD-1 is prevented, most typically by one in binding PD-L1 or PD-1 albumen in space The upper interaction for inhibiting the two.Therefore, the use of the inhibitor prevents the cancer cell for being loaded with PD-L1 to escape siberian crabapple System, and finally identified and removed by immune system.

Checkpoint inhibitor pembrolizumabIt is that being directed to for FDA approval is present in T cell The monoclonal antibody of PD-1 inhibition receptor.Verified Pembrolizumab is active to the cancer of three types: cannot cut Melanoma, unresectable kidney and the metastatic non-small cell lung cancer removed.It is expected that the above-mentioned three kinds of diseases in the U.S. in 2015 Disease incidence be respectively as follows: 73,870,61,550 and 188,000.Assumed mortality rate is respectively 9,940,14,080and 181,900 (SEER statistics).

For unresectable melanoma, usingOverall survival is 28% after treatment 60 months, Progression free survival rate is 18%.Although these data are significantly better than chemotherapy, be also greatly improved sky in terms of survival rate Between.

Brief summary of the invention

The present invention provides one kind and immunomodulator (the anti-CD3 immunotoxin as killed T cell) and checkpoint is used in combination Inhibitor, and cancer antigen is discharged during T cell Homeostatic proliferation new after existing T cell is depleted simultaneously, it is thin to kill cancer The method of born of the same parents.

It is an object of the invention to provide a kind of method for the treatment of cancer to subject in need, including give tested The anti-CD3 specific immunity toxin of person's sufficient amount is to exhaust the existing T cell of subject；To subject provide it is a kind of release by The therapy of the cancer antigen of cancer cell expression；And

A kind of checkpoint inhibitor of subject is given,

Wherein, in the presence of cancer antigen, the existing T cell of patient exhausts the regeneration and maturation for leading to new T cell. In some respects, anti-CD3 specific immunity toxin is A-dmDT390-bisFv (UCHT1).In some respects, checkpoint inhibits Agent is monoclonal antibody, inhibits the interaction of PD-1 and PD-L1.It can be the monoclonal antibody of anti-PD-L1, further, inspection Make an inventory of the monoclonal antibody that inhibitor may be anti-PD-1, such as pembrolizumab.In other respects, the release cancer The therapy of disease antigen refers to radiotherapy.Further, the step of giving checkpoint inhibitor is providing release cancer antigen After step.In other respects, provide release cancer antigen the step of the step of giving anti-CD3 specific immunity toxin it Afterwards；The step of giving checkpoint inhibitor is after providing the step of discharging cancer antigen.

In other respects, described the step of giving anti-CD3 specific immunity toxin be the 1st, 2,3,4 day for the treatment of to Give the anti-CD3 specific immunity toxin of multi-dose；Described the step of providing release cancer antigen, is carried out within the 5th day in treatment； Described the step of giving checkpoint inhibitor, is carried out within the 16th day in treatment, is carried out within every three weeks later primary.Further, the party Method includes the steps that repeating to provide release cancer antigen over time (such as at 1 day, 1 week, 1 month or 1 year).In some sides Face, it is thin that the cancer is selected from unresectable non-small cell lung cancer, unresectable hepatocellular carcinoma, unresectable neck squamous Born of the same parents' cancer, unresectable colorectal cancer, unresectable gastric cancer and unresectable melanoma.

The present invention also provides a kind of method for extending cancer patient's life span, the anti-CD3 including giving patient's sufficient amount Specific immunity toxin provides a kind of cancer antigen for discharging and being expressed by cancer cell to exhaust the existing T cell of patient, to patient Therapy, and, give a kind of checkpoint inhibitor of patient；Wherein, in the presence of cancer antigen, the existing T cell of patient The regeneration and maturation for leading to new T cell are exhausted, the life span of patient is thus extended.

The present invention also provides a kind of immune systems for preparing patient to identify and kill metastatic and/or relapse cancer cell Method, comprising: give the anti-CD3 specific immunity toxin of patient's sufficient amount to exhaust the existing T cell of patient, to patient A kind of therapy of cancer antigen for discharging and being expressed by cancer cell is provided, and gives a kind of checkpoint inhibitor of patient；Wherein, in cancer In the presence of disease antigen, the existing T cell of patient exhausts the regeneration and maturation for leading to new T cell, is thereby preparing for immune system To identify and kill metastatic and/or relapse cancer cell.

Other features and advantages of the invention by subsequent specification portion details, directly record by partial content specification , partial content is that implementation through the invention can obtain.The present invention will be special by institute in specification and claims The composition do not pointed out and method are realized and are obtained.

Detailed description of the invention

Fig. 1:After (A-dmDT390-bisFv (UCHT1)) treats 4 days (20 μ g/kg of accumulated dose), T cell Homeostatic proliferation causes the center CD8 memory T cells substantially to expand.Phase I clinical trial IB or the IIB stage, 8 CTCL by Examination person.The mSWAT value of the tumor load of all subjects changes within the scope of 14-212.All subjects show original T Cell is almost completely depleted, then new T cell Homeostatic proliferation.

Fig. 2: applicationThe clinical response for treating rear patient；

The amino acid sequence (SEQ ID NO:1) of Fig. 3: A-dmDT390-bisFv (UCHT1)；

Fig. 4: it is thin to kill mouse CD3 positive EL4T for the immunotoxin of anti-mouse CD3, the mouse analog of Resimmune Born of the same parents；

Fig. 5: anti-mouse CD3 immunotoxin inhibits the glioma induced by G1261mEGFRVIII cell transplantation Growth.

Detailed description of the invention

The present invention provides one kind and immunomodulator is used in combination (as killed T cell and be loaded with the cancer cell of CD3 antigen Anti- CD3 immunotoxin) and checkpoint inhibitor, and further associated release cancer antigen therapy (such as radiotherapy) kills cancer cell Method.For example, United States Patent (USP) 7,696,338 and 8,217,158 (Neville etc.) describes the immune poison of typical anti-CD3 Element.The life span of cancer patient is extended using this method, and is prevented and/or helped to kill metastatic or recurrent tumor And cancerous lesion.For example, patient kept without cancer state and/or life span extension at least six moon, 12 months, 24 months or longer. In some cases, patient is kept for indefinite duration without cancer.

Theoretical constraint is not considered, it is believed that when the anti-CD3 immunotoxin of T cell can be eliminated by giving cancer patient, No matter whether cancer has CD3, and immunotoxin all eliminates the T cell of existing normal, health expression CD3 antigen.This causes to suffer from New initial central memory T cells reactivity Homeostatic proliferation in person's body.Central memory-type cd8 t cell generally can not maintain to increase Grow state and Homeostatic proliferation signal.But in the presence of cancer antigen, during central memory-type cd8 t cell can be converted to Entreat memory effect form, make it possible it is this maintain.Because of this, it is very likely that new central memory-type cd8 t cell was breaking up Cheng Zhong, the cancer antigen sensitization being released, and then break up the effect type for becoming the cancer cell that can identify and kill its sensitivity Cd8 t cell.

According to certain aspects of the invention, new memory-type cd8 t cell occurs to the differentiation of CD8 effect type cell in cancer In the presence of disease antigen, the cancer antigen i) is released from the tumour cell of subject for example, by radiotherapy, And/or ii) from the tumor sample for example retained in the past；And/or iii) derive from synthetic antigen.No matter which kind of source, cancer Central memory-type cd8 t cell in the presence of disease antigen is divided into effect memory-type cd8 t cell and likely results in effect memory-type Cd8 t cell specific recognition and can kill all cells for being loaded with these antigens, such as existing cancer cell in subject's body, The cancer cell etc. that cancer return generates.

Before differentiation, therefore central memory-type cd8 t cell high level expression PD-1 simultaneously becomes the fine of checkpoint inhibitor Target spot.Joint inspection point inhibitor is likely to improve turn of the central memory-type cd8 t cell to effect memory-type cd8 t cell Change.The normal cell in addition, interaction of PD1/PD-L1 can disguise oneself as cancer cell, checkpoint inhibitor is by blocking example As the interaction of PD1/PD-L1 directly to help CD8 effector T cell to identify cancer cell.

In some respects, initial center memory-type cd8 t cell is exposed to cancer by providing radiotherapy to subject Antigen.Radiotherapy makes illuminated tumour cell release cancer antigen with its dead and decomposition in therapeutic scheme, helps It helps central memory T cells to be converted to effect type T cell, identify and kills the tumour cell for being loaded with the antigen of irradiation release.When Homeostatic proliferation is combined with cancer cell dissolution and antigen release, and antitumor immune response indefinite can exist.Therefore, new conversion Effect type T cell can immediately (existing tumour cell is present in the position that can not be irradiated) or in the future (such as tumour hair Raw transfer or recurrence) attack tumour cell.Therefore, anti-CD3 immunotoxin joint inspection point inhibitor, and selectable/usually Combination tumor Thrombosis (such as radiotherapy), prevent the permanent immunity of cancer return very for killing cancer cell and obtaining There may be synergistic effects.

" collaboration " or " collaboration " refer to that the interaction of three kinds of therapies is produced better than three kinds of therapies of exclusive use The combined effect of the sum of its effect.

Drug type and treatment method

The present invention, which provides, a kind of by following combination therapy to treat cancer and/or prevents the side of cancer metastasis and/or recurrence Method, the method includes 1) at least one immunomodulator, 2) at least one checkpoint inhibitor and 3) selectable, it is a kind of Cancer dissolution, antigen release therapy, such as radiotherapy.

" immunomodulator " of the present invention refers to improve, increase or supporting, promote the chemical combination of immune system activity Object.Especially, the immunomodulator improves the activity for fighting the immune system of one or more cancers.The published U.S. is special Benefit 20150166660 (full content of the patent refers in this patent as reference) describes immunotoxin molecule, quilt For the CD3 antigen to inhibiting tumor cell.Patent 20150166660 discloses these immunotoxins as immunomodulator with frightened The effect of people.Immunotoxin is most early in No. 7,696,338 authorized and No. 8,217,158 (Neville) United States Patent (USP) Middle record, the full content of this two patents refer in this patent as reference.It is used primarily for directly killing and is loaded with CD3 antigen The immunotoxin molecule of cancer cell be chimera or fusion protein, the recombinant toxin part comprising connecting antibody moiety is described Antibody moiety can specifically bind CD3 epitope.Antibody moiety is responsible for immunotoxin being integrated to tcr complex On CD3 ε γ subunit, enables the targeting of immunotoxin molecule specificity and be integrated in the T cell for being loaded with CD3 receptor.Once In conjunction with the toxin moiety of molecule will enter and kill cell.In some embodiments, the toxin moiety is, such as truncates Diphtheria toxin (DT) part or pseudomonas exotoxin A (ETA) toxin moiety, and the antibody moiety is anti-comprising AntiCD3 McAb Two scFv s of body.The amino acid sequence of anti-CD 3 immunotoxins A-dmDT390-bisFv (UCHT1) such as Fig. 3 and SEQ ID Shown in NO:1.The variant of the sequence also can be used, such as the piece of the variant of conservative replacement, proteolysis occurs for amino acid sequence Section includes and the variant not comprising methionine residues, the codon of optimization and/or humanization variants etc..In addition, serine egg (SEQ ID NO:2, the 191-198 referring to SEQ ID NO:1 are residual in such as Furin cleavage site RVRR:SVGS for white enzyme Base) or other possible sites cracking, without destroy cysteine 188-202 between disulfide bond.Any variant may It is used to treat or prevent cancer described herein, as long as variant retains immunotoxin activity.

The immunotoxin preparation of the present invention of bacterium is given, such as can be caused patient's body T is thin Born of the same parents group is depleted to the amount for being adequate to bring about immune system Homeostatic proliferation.T cell group is exhausted to be related to destroying or killing subject's body Inside at least 90-99% or more than (such as 100%) T cell, but may kill 50% or more in some cases (such as 55%, it 60%, 65%, 70%, 75%, 80% or 85%) is sufficient.

The checkpoint inhibitor being suitble in the method for the present invention includes but is not limited to target the preparation of PD-1 and target PD- The preparation of L1.Typical PD-1 preparation includes but is not limited to: PembrolizumabNivolumabPidilizumab etc..Typical PD-L1 preparation includes but is not limited to: AtezolizumabAvelumab(MSB001071BC), Durvalumab (Imfinzi^TM, MED14736), a kind of checkpoint AZD1775, Wee1 G2 serine/threonine protein kitase inhibitor, BMS-936559 (MDX-1105), MED14376, etc..In addition, in some respects, (another T is thin for the checkpoint inhibitor for targeting CTLA-4 Albumen on born of the same parents, its function are the buttons as one " closing ", guarantee that immune system is in normal condition, avoid siberian crabapple System excessively activation).Typical anti-CTLA-4 preparation includes but is not limited to: Ipilimumab

In some respects, antigen release therapy is provided in the combined therapy scheme, for example, giving a kind of dissolution of subject Cancer cell and the medicament or therapy that cancer antigen is discharged into the circulatory system.In some respects, antigen is released by radiotherapy It puts." radiotherapy " (XRT), which refers to using high-energy ray, to be reduced and kills cancer cell.Radiotherapy includes using X-ray, gal Horse ray and/or charged particle.Radiation (such as local radiation) can carry out in vitro (extracorporeal irradiation therapy) by instrument, Radioactive substance can be placed to cancer cell in vivo nearby to achieve the effect that radiotherapy (intracavitary irradiation therapy is also named short distance Radiotherapy).Alternatively, the total body irradiation of radioactive substance such as radioiodine can also be used.The type of radiotherapy Including but not limited to: stereotactic radiotherapy (SBRT), stereotaxis proton therapy (RSBPT) etc..

In other respects, antigen can also be by either along with the medicament of the energy dissolved cancer cell as immunotoxin comes Release.The medicament of this kind of dissolved cancer cell can be immunotoxin, for example, targeting the immunotoxin of certain types of cancer cell (such as the immunotoxin for targeting the HER-2 of breast cancer cell) or target the anti-of the neovasculature for supplying human tumor Body (such as antibody of anti-prostate-specific membrane antigen, anti-PMSA) etc..

Method

A kind of method that the present invention provides treating cancer and/or prevents cancer metastasis and/or recurrence, and then when extending existence Between, disease-free survival time and cancer return time.The method is related to giving a kind of combination treatment, comprising: I) at least one exempts from Epidemic disease regulator；II) at least one checkpoint inhibitor；And III) a kind of selectable, tumor lysis/antigen release therapy, example Such as radiotherapy.Usual this method include using means known in the art diagnosis subject with cancer, metastatic cancer and/or The step of relapsed cancer.

I. give anti-CD3 immunotoxin: after diagnosis, subject gives immunomodulator for example anti-CD3 immunotoxin first It induces reactive Homeostatic proliferation and improves the frequency (quantity, grade etc.) of the center (expansion) CD8 memory T cells.Due to treatment Effect even all will not significantly show in several years several weeks, some months after the treatment, therefore immediately begun to after diagnosing Immunological regulation be it is highly advantageous, it is initially desirable for making immunological regulation as early as possible.

The dosage of immunotoxin may be different according to understanding of those skilled in the art such as skilled medical practitioner to parameter And change.The dosage and medication of recommendation can determine in clinical test.Dosage is potentially based on following factor and changes: example The weight of such as patient, gender, the age, integral status, and/or disease type and stage, and whether given during administration Give other drugs etc..In general, the overall dosage of (plan carries out a period of time, such as 4 days) is big during a wheel chemotherapy About change in 5-60 μ g/kg weight range, for example, dosage is about 5,10,15,20,25,30,35,40,45,50,55 Or 60 μ g/kg weight.Specifically, giving the dosage of 20 μ g/kg weight in such as 4 days.0.5-5 μ g/kg weight is given in one day Dosage (the μ g/kg weight of about 0.5,1,1.5,2.0,2.5,3.0,3.5,4.0,4.5 or 5), be divided into 1-6 (such as 1,2,3,4,5 or 6) secondary administration, usually twice daily.The number of days or all numbers for treating progress can be according to the factors for influencing dosage and time And change (such as 2-10 days or longer are specifically 2,3,4,5,6,7,8,9 or 10 days).For example, the AntiCD3 McAb preparation A- given The dose of dmDT390-bisFv (UCHT1) is 20 μ g/kg, which shows as can induce Homeostatic proliferation (assuming that the anti-DT effect of patient Valence is lower than 22 μ g/ml).Within 4 day time, which is divided into 8 equal part dosage, gives an equal part dosage every time, gives daily Medicine twice, i.e. treatment in 1-4 days.A-dmDT390-bisFv (UCHT1) completes inducing T cell consumption usually in 14 days course for the treatment of To the greatest extent because anti-DT antibody titer can the 5th day increase so that influence subsequent T cell exhaust until potency reduce, such as 3 to 12 After month.To solve this problem, in some aspects, make immunotoxin therapeutic process using the truncated toxin sequence of disimmunity originality It extends to such as 5 or 6 days or one week or longer.

Normal T-cell numbers 2-3 Logarithmic degree is caused within 4 days to reduce (twice daily) specifically, giving anti-CD3 preparation for treating And induce reactive Homeostatic proliferation.T cell in the circulatory system is horizontal, including mature CD3T cell and central memory-type, effect Memory-type cd8 t cell is answered, can be detected by means known in the art come optimizing therapeutic regimen and obtains therapeutic purposes.Together Sample, other Immune System Components, such as it is directed to toxin moiety, other therapies or the ingredient, and/or tumour antigen of immunotoxin Antibody be considered as effective.In general, 15-38 days after administration, the regeneration of T cell improve the center CD8 memory T cells Ratio, be administration before 10-20 times.The marker CD45RA of these cells expression activation^-/low, CD3, CD27and CD8.

II. tumour antigen discharges.Give anti-CD-3 preparation and other release cancer antigens therapy groups be combined into proliferation T it is thin Born of the same parents become " being trained to " T cell and provide chance.Therefore, the method for the present invention further include killed in a manner of release tumor antigen it is swollen The step of oncocyte, or the step of exposing cancer antigen to developmental central memory T cells is to promote to generate for this The immunological memory cell of kind cancer antigen.In short, the 5-6 days tumor-cell antigens after giving anti-CD 3 immunotoxins apply one It plants or a variety of suitable preparation or therapy are released or other modes give subject.Used antigen release therapy should not damage Evil lymph structure and function.

If oncolysis therapy is local irradiation, implements dosage and determined by radiation oncology doctor, specifically existed Single therapy (1-2 days i.e. after giving anti-CD3 immunotoxin treatment in 4 days) are given within 5th day, dosage is changed in 3-20Gy, such as 3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19 or 20Gy.To consider when selecting dosage it is many-sided because Whether element, including patient receive chemotherapy, if there is secondary morbidity to become, operation consent carries out radiotherapy after still perform the operation, and perform the operation at Power etc..Large Dose Irradiation (such as single is within the scope of 10-20Gy) may improve the reactivity of the lesion other than irradiation area, and Therefore more significant effect is generated to immunological regulation.

As previously mentioned, radiotherapy can also be replaced using other antigen release therapies or implemented together with radiotherapy.For example, if Cancer cell is loaded with CD3 antigen, then anti-CD3 preparation unexpectedly can equally kill cancer cell and released antigen.

Other kinds of antigen can such as be taken from the tumour of subject or the antigen preparation of biopsy sample.The sample can To be saved, the method that a portion describes at reasonable time interval with the present invention is combined, and such as gives 4 days anti-CD3 systems 1-2 days after agent.

The step of discharging cancer antigen carries out early stage treatment, so that immune system is enough to supervise rapidly after recurrence Control, identification and elimination novel tumor are without further treating.However, in other respects, in order to further enhance immune response, Antigen release therapy can be applied again over the course for the treatment of, be similar to immunization scheme.If cancer return, can be easy Ground is found, because can execute the treatment of released antigen at that time.However, if not finding or detecting tumour cell again Raw, it may be by being administered tumour cell or containing the segment enhancement effect from initial tumor antigen, the initial tumor Save thus.In this case, cell or segment can be in initial treatments for example after 3-6 months, if it is desired, can also With and/or longer interval (such as several years) be administered afterwards, using as " reinforcing agent ".It is by improving the continual immune prison of patient Superintend and direct and with the natural ability that tumour is struggled, promote to non-irradiated and/or shifting and/or the cancer that recurs is controlled It treats.The exposure dose and scheme are the same as the first treatment above.

III. checkpoint inhibitor is given

After antigen release therapy, such as after about 7-10 days, at the 16th day for the treatment of, checkpoint inhibitor is given (1-4 days for the treatment of are the time for giving anti-CD3 immunotoxin).Hereafter, a checkpoint inhibitor is about given every 3 weeks, is held Continuous some months (such as 1-6 months) and/or until until subject's vivo detection is less than cancer cell, or until checkpoint inhibitor Until the side effect of generation causes to deactivate checkpoint inhibitor.

The dosage of checkpoint inhibitor is referring to FDA being approved range described in new drug or FDA to trial drug according to approval Test new drug Policy Conditions range.On the one hand, due to the association of use in conjunction anti-CD3 immunotoxin and checkpoint inhibitor The influence of same-action, I phase clinical medicine be treated in combination the stage, the dosage of checkpoint inhibitor relative to clinical indication It reduces.For example, one or more of checkpoint inhibitor are according to such as about 0.1-100mg/kg or about 1-75mg/kg or big The dosage of about 5-50mg/kg is given.Usually the 4th day for giving anti-CD3 preparation or checkpoint inhibitor is given in 4 days.For up to To desired effect, can primary enough administrations, or be divided into low dose, and/or by week, moon repeat administration.

In the entire life process of patient, therapeutic process can be repeated as desired for carrying out, especially cancer return when.But It is, for the repetitive treatment, to be not usually required to repeat to give anti-CD3 immunomodulator, be given only local tumor radiotherapy Checkpoint inhibitor is combined or be not given in combination to (as previously described).

The method of the present invention is by giving including the active ingredient (sometimes encoding their nucleic acid sequence) and pharmacology The composition of suitable (physiology is compatible) carrier is gone up to implement.The preparation of the composition is well known to those skilled in the art. Specifically, the composition is prepared into liquid preparation or suspending agent.The active constituent can with it is pharmaceutically acceptable and with work Property mixed at the compatible excipient of split-phase.Suitable excipient is, for example, water, salting liquid, glucose, glycerol, ethyl alcohol etc., or Its composition.In addition, the composition may include a small amount of auxiliary material, such as wetting agent or emulsifier, pH buffer etc..

In general, anti-CD3 preparation and checkpoint inhibitor (and/or antigen releasing agent, if the latter is not radiation) Independent component as modality of cancer treatment is administered together (joint).Likewise, any preparation usually as individual component into Row administration.Be administered simultaneously or be not administered simultaneously can make receive treat subject blood flow in there are one or more preparations. But it is carried out before antigen release therapy is preferably bred after T cell group is reduced or again or in reproductive process.Usually exist Memory T cells breed again and give checkpoint inhibitor in conversion process, and expand effect type T cell activity.Administration is to be System property.

Although preparation of the present invention is administered usually as independent component, not such was the case with for situation.Comprising a kind of or The composition of a variety of anti-CD3 preparations and the/one or more checkpoint inhibitor of shield and/or one or more antigen releasing agents It is included therein.

The medication of chemotherapeutant familiar to those skilled in the art, the composition (preparation) may be by much Any one in the suitable way known is administered, including but not limited to: injection, sucking, oral, intravaginal administration, nasal cavity Administration, external application, eye drops, spraying, intra-tumoral injection, the transplanting of composition or device for discharging medicament etc..It is common, administration Mode is intravenous injection or subcutaneous injection.

In addition, the composition can be combined with other treatment form, for example promote substance (such as tubercle bacillus of immune system Or part thereof), it is various other antitumor/anticancer chemotherapy agent (such as platinum medicine cis-platinum, amethopterin), anodyne, anti- Dizziness medicine, antiallergic (such as antihistamine), nutritional supplement, antidepressants.Equally, operation, which can also be used as, entirely treats A part, such as any suitable one or more tumours of time excision before or after the method for the present invention starts.Other Associable cancer therapy will be described in detail below.

The preparation that the present invention describes can any required time after cancer is made a definite diagnosis according to any suitable therapeutic scheme or Timetable is administered.It can be carried out before, after or at the same time in other anticancer drugs or therapy.For example, it can be at it Before his cell toxicity medicament or therapy or operative treatment start and/or simultaneously or after (such as a few days or a few weeks) be administered.If It is administered with other treatment form " together ", it can be within the successively shorter time, such as with list in a few minutes, a few houres or several days Only composition administration, or use include at least one (i.e. one or more kinds of) drug (i.e. anti-CD3 preparations, checkpoint suppression Preparation or antigen release therapy, if not radiotherapy) and one or more kinds of anticancer drug single composition.

The cancer types for the treatment of

In some respects, it is considered as suitable for using the patient that the method for the present invention is treated being that those are diagnosed with cancer cell and do not have There is the cancer patient of surface C D3 epitope, i.e. cancer cell surfaces do not have (missing) CD3 epitope.In this aspect, if anti-CD3 is immune Toxin is used as immunomodulator, then it plays effect mainly as immunomodulator, does not show directly to kill cancer cell Effect.Cancer without surface C D3 epitope includes that any non-T cell leukaemia or lymph cancer are (i.e. any to be not belonging to T cell The cancer of leukaemia or lymph cancer)；Acute myeloid leukaemia (AMK)；Adrenocortical carcinoma；Atypia deformity/rod-shaped tumour；In Pivot nervous system cancer；Basal-cell carcinoma (such as non-black melanoma)；Cholangiocarcinoma；Bladder cancer；The outer bladder cancer of liver；Osteocarcinoma (such as Juventus Tumour, osteosarcoma and the malignant fibrous histiocytoma of sarcoma family)；Brain stem glioma；Brain tumor (such as astrocytoma, brain It is thin with spinaloma, central nervous system atypia deformity/rod-shaped tumor, central nervous system plumule tumor, central nervous system reproduction Born of the same parents' tumor etc.)；Craniopharyngioma；Ependymoma；Breast cancer；Tumor of bronchus；Burkitt lymphoma；Gastroenteric tumor；Cardiac tumor； Cervical carcinoma；Chordome；Chronic lymphocytic leukemia (CLL)；Chronic granulocytic leukemia (CML)；Chronic myeloproliferative Disease；Colon cancer；The carcinoma of the rectum；Craniopharyhgeal canal cancer；Cutaneous T-cell lymph cancer；Tumors of Extra-hepatic Bile Duct；In situ ductal carcinoma (DCIS)；Embryo Bud tumour；Carcinoma of endometrium；The cancer of the esophagus；Esthesioneuroblastoma；Ewing sarcoma；Germinoma outside cranium；Sexual gland is external Cell colonization tumor；Cancer eye (intraocular melanoma, retinoblastoma)；Bone fibrous histiocytoma；Osteosarcoma, gallbladder cancer；Stomach Cancer；Gastrointestinal associated cancers tumour；Gastrointestinal stromal tumor (GIST)；Pregnant trophoblastic tumor；Glioma；Hairy cell leukemia；Head Neck cancer；Heart cancer；Liver cancer；Hypopharyngeal cancer；Intraocular melanoma；Islet-cell tumour；Pancreatic Neuroendocrine Tumors；Kidney (example Such as nephrocyte and the nephroblastoma)；Langerhans cell histiocytosis；Laryngocarcinoma；Leukaemia；Liver cancer (primary)；Leaflet Carcinoma in situ (LCIS)；Lung cancer (non-small cell, cellule)；Lymthoma；Macroglobulinemia Waldenstron；Male mammary gland Cancer；Malignant mesothelioma；The cervical metastasis squamous cell carcinoma of primary tumor concealment is related to the main middle harness cancer of NUT gene； Carcinoma of mouth；Endocrine neoplasia syndrome；Myelodysplastic syndrome；Myeloproliferative disorder/marrow and outside myeloproliferative tumour； Chronic granulocytic leukemia (CML)；Acute myeloid leukaemia (AML)；Huppert's disease；Chronic myeloproliferative imbalance； Nasal cavity or nasal sinus cancer；Nasopharyngeal carcinoma；Neuroblastoma；Non-Hodgkin lymphoma；Carcinoma of mouth；Carcinoma of mouth (oral cavity)；Mouth Lip and oropharyngeal cancer；Osteosarcoma and malignant fibrous histiocytoma of bone；Oophoroma；Cancer of pancreas；Pancreatic Neuroendocrine Tumors (pancreas islet Cytoma)；Papillomatosis；Chromaffionoma；Thyroid cancer；Carcinoma of penis；Pharynx cancer；neochromocytoma；Hypophysoma；Slurry is thin Palpebral edema tumor/Huppert's disease；Pleuropulinonary blastoma；CNS lymthoma；Prostate cancer；The carcinoma of the rectum；Nephrocyte (kidney) cancer； Salivary-gland carcinoma；Sarcoma (Juventus, Ka Boji, osteosarcoma, rhabdomyosarcoma, soft tissue, uterus)；Cutaneum carcinoma (melanoma, Merkel Cell cancer, non-black melanoma)；Small Cell Lung Cancer, carcinoma of small intestine, squamous cell carcinoma；The squamous neck cancer of primary tumor concealment；Transfer Property gastric cancer；Carcinoma of testis；Laryngocarcinoma；Thymoma and thymic carcinoma；Thyroid cancer；Renal plevis and ureter transitional cell cancer；Trophoderm is swollen Tumor, it is gravidic；Urethral neoplasms；Carcinoma of endometrium；Sarcoma of uterus；Carcinoma of vagina；Carcinoma of vulva；Waldenstrom macroglobulinemia Disease；The nephroblastoma；Nasal cavity and nasal sinus cancer；Nasopharyngeal carcinoma；Neuroblastoma；Non-small cell lung cancer, cutaneous melanoma, Huo Qi Golden lymthoma and their transfer and recurrence.

Other combination treatments

In some respects, other toxin formulations and/or other therapies can also be used to realize in the treatment for example to kill entirely Portion's cancer cell causes the purpose of tumor regression, such as a short-term gamma therapy.Therefore, it can give one or more other anti- Cancer preparation or anti-cancer modalities or therapy, for example including but be not limited to: target specific tumour or generate tumour vascular cell Malicious immunotoxin, cell toxicant anti-tumor drug such as alkylating agent cis-platinum, carboplatin, oroxaliplatin；Disguise as purine The antimetabolite of (such as imuran, mercaptopurine) or pyrimidine；Plant alkaloid and terpene, such as vinca alkaloids such as Changchun New alkali, vinblastine, vinorelbine, eldisine；Podophyllinic acid lactone, Etoposide and Teniposide；Taxanes such as Japanese yew Alcohol；Topoisomerase enzyme inhibitor I type includes camptothecine, Irinotecan and topotecan, and topoisomerase enzyme inhibitor II type is for example Amphidine, Etoposide, etoposide phosphate and Teniposide；With cytotoxic antibiotics for example D actinomycin D, anthracycline, It is adriamycin, daunomycin, soft than star, idarubicin, epirubicin, bleomycin, plicamycin, mitomycin；Gene therapy (such as the nucleotide delivery of anticancer agent will be encoded to tumour), cancer surgery/resection；Hormonotherapy；Angiogenesis suppression Preparation administration；Other immunomodulators or treatment (such as allogene/homologous hematopoietic stem cell transplantation) is administered；By external beam radiotherapy The radiotherapy for treating (EBRT)；Or the internal therapentics by brachytherapy, electrochemotherapy；Ultraviolet light (UV) treatment Deng.

Following embodiment is intended to illustrate the various representative aspects of invention, but is not necessarily to be construed as to of the invention Limitation.

Specific embodiment

Embodiment 1A-dmDT390-bisFv (UCHT1) fusion proteinCombined with Ionizing Radiation and Anti- PD-L1 treats unresectable non-small cell lung cancer, unresectable liver cancer, unresectable Head and neck squamous cell carcinoma, no The adaptability clinical trial design of resectable colorectal cancer, unresectable gastric cancer and unresectable melanoma.

It includes to one or more particular aspects of researching and designing and based on tested that adaptability clinical trial design, which is one, The hypothesis of person's data analysis (usually ephemeral data) plans the research improved as expected.It is anticipated that the time point of plan It carries out, with double blind or non-blind mode, the analysis with formal statistical hypothesis testing to the data of accumulation.

This experiment carries out dosage first and increases research to assess safety of the conjoint therapy to various cancers.From any pre- All enrolled subjects for the unitized dose level first planned observe its DLT in 28 days of first time administration.Once plan The number of subjects (preceding 3 subjects in 3+3 design) of any dosage group passes through 28 days DLT windows, then the dosage level quilt It removes, studies and carried out to subsequent dose level or next stage.Since target is detection safety, and non-validity, therefore can To include a variety of unmet tumor types and be not necessarily to can be carried out subsequent dose level until the point that works, so as to study It faster registers and is carried out as early as possible to next step.The Dose Escalation of anti-PD-L1 has reached 1mg/kg in phase iii clinical trial, and The dosage (3 subjects in 3+3 design) of 2.5mg/kg and final 5mg/kg will be developed to.The dosage of Resimmune will be protected It holds in constant 20 μ g/kg, every 4 days primary, twice daily, because the dosage can induce T cell and exhaust and generate therefrom The increase of Homeostatic proliferation and subsequent similar 20 times of cell of Tcm (central memory T cells).Radiological dose, preferably SBRT, by Radiation oncology doctor selects within the scope of 14-24Gy according to the size of metastatic carcinoma, it is intended to alleviate and it is non-effective.Have in needs assessment In the Dose Escalation stage of efficiency, we can choose addition and remove the second test group of Resimmune to determine the use of The increased degree of validity of Resimmune combination.Due to that can obtain data, we will be predicted most using statistical method Good case queue size.

We predict that Resimmune combines anti-PD-L1 relative to the minimum raising 135% of the anti-PD-L1 effect of exclusive use And it is possible to be increased to 250%.This raising is likely due to combination treatment can improve tumour antigen in tumor environment And eliminate T cell suppression mechanism.

Exclude: the patient with hepatocellular carcinoma shows as not having duplication hepatovirus；There is the patient of history of heart disease to need to exclude. Referring to researcher's handbook: treating Alibert's disease using A-dmDT390-bisFv (UCHT1).

For unresectable melanoma, the anti-checkpoint PD-1 inhibitor is being used60 after treatment A month overall survival is 28%, and Progression free survival rate is 18%.In next step the challenge for the treatment of be to find out why certain patients There is response and some patients do not have, to improve response rate and overall survival.

Clinical research has shown that, using checkpoint inhibitor for treating cancer, quantity and the clinic of tumour somatic mutation have Efficiency it is very related, even if the lymphocyte (TILs) of infiltration tumour can be identified using lower somatic mutation quantity it is swollen Tumor.The count issue of somatic mutation leads to few TILs or the insufficient TILs of T cell activity.In mouse tumor research, The preparation for inducing Homeostatic proliferation improves the anticancer activity of adoptive transfer T cell.PD-1/PD-L1 is blocked to improve central note The effector function for recalling type T cell may be because of central memory T cells high level expression PD-1.

Skin T cell lymphoma patient's body T cell is induced to exhaust and subsequent central T cell Stable state breeding, causes to recycle central memory T cells and increased at the 16th day to 140 days (see Fig. 1, being demonstrated by 60 days data) It is 20 times long.These central memory T cells can be converted into effect memory T cells, 20 times of growth in the presence of tumour antigen So that the growth of tumour-specific center memory T cells quantity increased and therefore effect type T cell is caused to convert, causes to exempt from The tumor suppression that epidemic disease is adjusted.This should be applicationTherapy causes to promise completely for a long time immunoregulatory Mechanism, it is undiscovered in traditional treatment (see Fig. 2).

Checkpoint inhibitor helps to block PD-1/PD-L1 in this process.The therapeutic scheme to melanoma patients especially Effectively, because melanoma has high-caliber somatic mutation.However, cancer of the therapeutic scheme for other low somatic mutations Disease is also effective, such as hepatocellular carcinoma, be because promote to the compensation of central memory T cells quantity (by Undertake), central memory T cells can be converted into effect type T cell (by the exposure of antigen caused by radiotherapy).

In illustrative aspect, using anti-CD3 preparation RESIMMUNE^TM, anti-PD-L1 and palliative radiotherapy three combination come Treatment suffers from the patient of unresectable IV phase cancer.The effect of palliative radiotherapy be help discharge melanoma-associated antigen to Cause dendritic cells and T cell to infiltrate and (is shown in Table 1) into tumour.

Table 1Combine the design scheme example and timetable of anti-PD-L1 and radiotherapy

Second target is to be treated with the high-throughput deep sequencing technology detection area people TCR V- β to be preferably characterized in The amplification and clone's characteristic in journey periphery T cell library.The variation in T cell library caused by immunotherapy and peripheral T cell Homeostatic proliferation/ Percentage/phenotype (modulating T cell and IL-17+T cell) of activation and immunosuppressant cell is related.These change with individually The nothing using anti-PD-L1 or anti-PD-1 treatmentThe reaction obtained with radiotherapy compares, and can demonstrate,prove The mechanism of bright combination treatment.In the case where main stimulator antigen is unknown, feature and efficacy of drugs that combination periphery TCR is cloned Biomarker is the valuable method for tracking the tumor specific T cells in peripheral blood, can be used as the biomarker of omen Object is to evaluate reaction or resistance of the melanoma patients to immunotherapy.

2 mouse model of embodiment prove anti-CD 3 immunotoxins to the effect of tumour growth, with study checkpoint inhibitor and The synergistic effect of anti-CD 3 immunotoxins

In order to prove the antitumous effect of anti-CD 3 immunotoxins, we have developed be similar to anti-human CD3 immunotoxin The immune poison of the anti-mouse CD3 of Resimmune.Anti-mouse CD3 immunotoxin has structure identical with mankind's counterpart and same Sample has specificity to the epsilon chain of CD3 complex.Immunotoxin can effectively kill the mouse CD3 positive T of in vitro culture Cell (see Fig. 4).

In solid tumor models, we used mouse C57BL/C with normal immunological function and modified small Rat glicoma cells system Gl261mEGFRvIII.The cell line is overexpressed mouse EGFRvIII, but does not express CD3, therefore fights Mouse CD3 immunotoxin is insensitive.Gl261mEGFRvIII cell (5X10⁵/ mouse) it is inoculated to C57BL/C mouse Right side.When Growth of Tumors Transplanted to 150mm³, by tail vein injection, give experimental group anti-mouse CD3 immunotoxin, every note The PBS solution (40 μ g/kg) of 0.1ml anti-mouse CD3 immunotoxin is penetrated, injection is primary daily, and totally 4 days.Every mouse of control group Give the PBS buffer solution of 0.1ml.As shown in figure 5, anti-mouse CD3 immunotoxin can significantly delay tumour growth.

Since anti-mouse CD3 immunotoxin is not directly placed on Gl261mEGFRVIII cell, the life of cell will not influence It is long, therefore as seen in Resimmune-CTCL clinical test, the inhibiting effect of Gl261mEGFRvIII transplantation tumor is then led To depend on its immunoloregulation function.It is considered that anti-CD3 immunotoxin there is anti-tumor activity to be because it can cause T thin Born of the same parents, which exhaust, causes cd8 t cell by Homeostatic proliferation rapid amplifying, kills tumour cell.Currently known tumor cells expression inspection Molecule (such as PD-L1) is put to escape the attack of T cell, therefore, if we predict that anti-CD 3 immunotoxins and inspection are used in combination Point inhibitor (such as anti-PD-1 or anti-PD-L1 antibody), will reach more far-reaching in mouse tumor model and treating human cancer With lasting tumor inhibitory effect.

It is contemplated that anti-mouse CD3 immunotoxin and checkpoint inhibitor is used in combination, (such as anti-mouse PD-1 or anti-is small Mouse PD-L1 antibody), total response rate can at least reach the 135% of non-treatment control group；Tumor inhibitory effect (or tumor regression Effect) it is likely to be breached the 250% of non-treatment control group.In mouse tumor model, with continuous 4 days tail vein injections or it can swell The method of anti-EGFR VIII immunotoxin of the intratumor injection based on diphtheria toxin (DT) is come release tumor antigen

Although preferred embodiment has been described in the present invention, it will be appreciated by the appropriately skilled person that the present invention can be Implement after modifying in the spirit and scope of claims.Correspondingly, the present invention should not necessarily be limited by above-described embodiment, but into One step, it should comprising whole modifications in the spirit and scope mentioned herein with this specification and its be equal.

Sequence table

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Claims (16)

1. a kind of method to subject's cancer in need, which is characterized in that the described method includes:

The anti-CD3 specific immunity toxin for giving subject's sufficient amount exhausts the existing T cell of subject,

A kind for the treatment of method for discharging cancer antigen is provided to subject, the antigen is expressed by cancer cell, and

A kind of checkpoint inhibitor of subject is given,

Wherein, in the presence of cancer antigen, exhausting the existing T cell of subject leads to T cell new in subject's body Regeneration and maturation.

2. the method as described in claim 1, which is characterized in that the anti-CD3 specific immunity toxin is A-dmDT390- bisFv(UCHT1)。

3. the method as described in claim 1, which is characterized in that the checkpoint inhibitor inhibition PD-1's and PD-L1 is mutual Effect.

4. method as claimed in claim 3, which is characterized in that the checkpoint inhibitor be anti-PD-L1 monoclonal antibody or Anti- PD-1 monoclonal antibody.

5. the method as described in claim 1, which is characterized in that the treatment of the release cancer antigen refers to radiotherapy.

6. the method as described in claim 1, which is characterized in that described the step of giving checkpoint inhibitor is antitumor in release It is executed after the step of antigen.

7. the method as described in claim 1, which is characterized in that give anti-CD3 special the step of the release antitumor antigens It is executed after the step of specific immunological toxin；With, the step of giving checkpoint inhibitor the step of discharging antitumor antigens it After execute.

8. the method as described in claim 1, which is characterized in that described to give anti-CD3 specific immunity toxin for the treatment of 1, give multi-dose anti-CD3 specific immunity toxin within 2,3,4 days；

The step of discharging antitumor antigens executes on the 5th day treatment；

The step of giving checkpoint inhibitor executes on the 16th day treatment, executes every 3 weeks later primary.

9. the method as described in claim 1, which is characterized in that further comprise repeating release anti-tumor over time Former step.

10. the method as described in claim 1, which is characterized in that the cancer is selected from unresectable non-small cell lung cancer, no Resectable hepatocellular carcinoma, unresectable Head and neck squamous cell carcinoma, unresectable gastric cancer, unresectable colorectal cancer With unresectable melanoma.

11. a kind of method for extending tumor patient life span, which is characterized in that the described method includes:

The anti-CD-3 specific immunity toxin for giving patient's sufficient amount removes the existing T cell of patient's body,

A kind for the treatment of method for discharging cancer antigen is provided to patient, the antigen is expressed by cancer, and

A kind of checkpoint inhibitor of patient is given,

Wherein, in the presence of cancer antigen, exhausting the existing T cell of subject leads to T cell new in subject's body Regeneration and maturation, to extend the life span of patient.

12. a kind of immune system by preparing patient is suffered from identifying and killing metastatic and/or relapsed cancer to improve The method of metastatic and the overall response rate of/relapsed cancer subject, which is characterized in that the described method includes:

The anti-CD3 specific immunity toxin for giving patient's sufficient amount exhausts the existing T cell of patient,

A kind of checkpoint inhibitor of patient is given, and

A kind of radiotherapy for metastatic lesion is provided to patient to discharge cancer antigen, the antigen is expressed by cancer,

Wherein, in the presence of cancer antigen, exhausting the existing T cell of subject leads to T cell new in subject's body Regeneration and maturation, so that preparing a kind of immune system identifies and kills metastatic and/relapsed cancer, relative to exclusive use Checkpoint inhibitor (without the use of anti-CD3 specific immunity toxin and radiotherapy) improves at least 135%.

13. a kind of kit, which is characterized in that the kit includes that anti-CD3 specific immunity toxin and checkpoint inhibit Agent.

14. kit as claimed in claim 13, which is characterized in that the anti-CD3 specific immunity toxin is A- dmDT390-bisFv(UCHT1)。

15. kit as claimed in claim 13, which is characterized in that the checkpoint inhibitor inhibits PD-1's and PD-L1 Interaction.

16. kit as claimed in claim 15, which is characterized in that the checkpoint inhibitor is that anti-PD-L1 monoclonal is anti- Body or anti-PD-1 monoclonal antibody.