Patent Application: US-201314654648-A

Abstract:
the present disclosure provides agents for treating and / or preventing resistance of tumor cells to radiation therapy , the use and relevant method thereof .

Description:
as used herein , the term “ agent capable of inhibiting b7 - h1 / pd1 signaling ” is used in its broadest sense to include any agents that can reduce or inhibit b7 - h1 / pd1 signaling , include , but not limited to , agents that inhibit the transcription , translation and modification of genes encoding b7 - h1 or pd1 , agents that influence the activity of b7 - h1 or pd1 protein , or agents that otherwise influence the direct or indirect interaction of b7 - h1 with pd1 . in certain embodiments of the present disclosure , the agent can be an agent capable of inhibiting the activity of b7 - h1 or pd1 protein , for example , a blocking antibody to b7 - h1 or pd1 . in a specific embodiment , the blocking antibody can be a specific monoclonal antibody , polyclonal antibody , humanized antibody , chimeric antibody or an antigen - specific fragment ( e . g ., fab , fv , scfv antibody fragment , or the like ) thereof . a person skilled in the art will appreciate that any agent able to specifically bind to b7 - h1 or pd1 protein and influence the function and / or structure thereof may potentially act as the “ agent capable of inhibiting b7 - h1 / pd1 signaling ” of the present disclosure . as used herein , the term “ radiation therapy ” or “ radiation treatment ” includes , for example , fractionated radiation therapy , non - fractionated radiation therapy and super - fractionated radiation therapy , as well as a combination of radiation and chemotherapy . the type of radiation may further include ionizing ( γ ) radiation , particle radiation , low energy transfer ( let ), high energy transfer ( het ), x - ray radiation , uv radiation , infrared radiation , visible light , photosensitizing radiation , etc . in one embodiment of the present disclosure , the radiation therapy or radiation treatment is a single or multiple x - ray irradiations , e . g ., 1 , 2 , 3 , 4 , 5 , 6 or more x - ray irradiations . preferably , the dosage of x - ray used in each irradiation may be 5 - 20 gy , such as 5 - 8 , 5 - 12 or 5 - 15 gy . in particular , when multiple x - ray irradiations are administered , the interval between irradiations may be one to several hours ( e . g ., 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 or up to 24 hours ), one to several days ( e . g ., 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 days , etc . ), or even one to several months ( e . g ., 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10 , 11 or 12 months ). as used herein , the terms “ chemotherapeutic agent ”, “ chemical therapeutic agent ” and “ agent for chemotherapy ” can be used interchangeably . it includes a composition comprising a single active ingredient or a combination of multiple chemotherapeutic agents . in a subject in need of a therapy , chemotherapy can be used in combination with surgery or radiation therapy , or with other forms of anti - tumor therapies , for example , the “ agent capable inhibiting b7 - h1 / pd1 signaling ” of the present disclosure . in particular , the chemotherapeutic agent includes , but not limited to , adriamycin , cyclophosphamide and taxanes [ paclitaxel ( taxol ) and docetaxel ( taxotere )], capecitabine ( xeloda ), gemcitabine ( gemzar ), vinorelbine ( navelbine ), tamoxifen , aromatase inhibitors ( arimidex , femara , aromasin ), 5 - fu withfolinic acid , irinotecan ( camptosar ), oxaliplatin , cisplatin , carboplatin , estramustine , mitoxantrone ( novantrone ), prednisone , vincristine ( oncovin ), etc ., or a combination thereof . in the context of the present disclosure , the term “ resistance to radiation therapy ” means that cells ( e . g ., tumor cells ) do not lose their ability to continue to propagate and / or grow after being through radiation therapy or treatment . generally , such resistance can lead to decrease or loss of the efficacy of the radiation therapy in tumor treatment , which in turn can cause tumor recurrence . in the context of the present disclosure , the term “ tumor ”, “ cancer ” or “ hyper proliferative disease ” refers to the growth and proliferation of any malignant or benign cancerous cells , including all transformed cells and tissues and all cancerous cells and tissues . examples of the cancer include , but not limited to , carcinoma , lymphoma , blastocytoma , sarcoma , leukemia or malignant lymphatic tumors . specific examples of the cancer include squamous cell cancer ( e . g ., epithelium squamous cell cancer ), lung cancer , including small cell lung cancer , non - small cell lung cancer , lung adenocarcinoma and squamous cell cancer of the lung , peritoneal cancer , hepatocellular cancer , gastric cancer , including gastrointestinal cancer , pancreatic cancer , glioblastoma , cervical cancer , ovarian cancer , liver cancer , bladder cancer , hepatoma , breast cancer , colon cancer , rectal cancer , colorectal cancer , endometrial cancer or uterine cancer , salivary gland cancer , kidney cancer , prostate cancer , vulvar cancer , thyroid cancer , liver cancer , anus cancer , penis cancer , and head and neck cancer . further examples of the cancer are listed in the elsewhere of the present disclosure . in particular , the tumor is selected from : breast cancer , ovarian cancer , bladder cancer , lung cancer , prostate cancer , pancreatic cancer , colon cancer and melanoma and / or cells thereof , for example , breast cancer cells ( such as tubo cells ) or myc - cap tumor cell line . in some embodiments of the present disclosure , the agent capable of inhibiting b7 - h1 / pd1 signaling of the present disclosure or the composition comprising the agent is administered within several hours ( e . g ., 10 - 48 hours ), several days ( e . g ., 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 or 9 days ) or several weeks ( e . g ., 1 - 8 , 1 - 7 , 1 - 6 , 1 - 5 , 1 - 4 , 1 - 3 or 1 - 2 weeks ) after radiation therapy or radiation treatment . when desired , the agent or the composition can be administered for multiple times , for example , 2 , 3 , 4 , 5 , 6 or more times , and the interval between administrations can be several hours , one day , several days ( e . g ., 2 - 30 , 2 - 25 , 2 - 20 , 2 - 15 , 2 - 14 , 2 - 13 , 2 - 12 , 2 - 11 , 2 - 10 , 2 - 9 , 2 - 8 , 2 - 7 , 2 - 6 , 2 - 5 , 2 - 4 , or 2 - 3 days ), one to several months or longer . for the prevention or treatment of resistance to radiation therapy , the dosage and manner for administrating the agent or composition of the present disclosure can be decided by physicians according to criteria known in the art . the administered concentration and dosage of the agent that inhibits b7 - h1 / pd1 signaling may depend on the type of the cancer to be treated , the severity and course of the disease , the size of the tumor , the degree of metastasis , the aim of administration is preventive or therapeutic , previous therapy , the patient &# 39 ; s medical history and response to antibodies , and the discretion of the attending physician . for repeated administration over several days or longer , depending on the condition , the treatment can be maintained until a desired suppression of the symptoms is achieved , for example , a reduction of tumor size / volume and reduction of metastasis . the course of treatment can be monitored by a conventional method or analysis based on the criteria known to physicians or any person skilled in the art . specifically , for an antibody , the dosage to be administered may range from 0 . 1 to 100 mg / kg of patient &# 39 ; s body weight , e . g ., 0 . 1 to 20 mg / kg of patient &# 39 ; s body weight , or 0 . 1 to 10 mg / kg of patient &# 39 ; s body weight . in general , human antibodies have longer half - life in humans than antibodies derived from other species due to immune responses to foreign polypeptides . therefore , a lower dosage of a human antibody and a lower frequency of administration are generally possible . furthermore , the dosage and frequency of administration of an antibody can be reduced by enhancing the uptake and the tissue penetration ( e . g ., into the brain ) of the antibody through a modification such as lipidation . the pharmaceutical composition according to the present disclosure can comprise a pharmaceutically acceptable excipient , carrier , buffering agent , stabilizer or other materials known to those skilled in the art . such materials shall be non - toxic and shall not interfere with the efficacy of the active ingredient . such materials may include any solvent , dispersion media , coating , antibacterial and antifungal agent , isotonic and absorption delay agent , physiologically compatible substance , etc . the pharmaceutically acceptable carrier can be , for example , water , saline , phosphate buffered saline , glucose , glycerol , ethanol or the like , and a combination thereof . in many cases , the pharmaceutical composition may comprise an isotonic agent , for example , sugar , a polyol such as mannitol , sorbitol , or preferably sodium chloride . the pharmaceutically acceptable substance can also be a wetting agent or a small amount of auxiliary substance , such as a moisturizer or an emulsifying agent , a preservative or a buffering agent , which can increase the shelf life or efficacy of the antibody . the concrete properties of the carrier or other materials will depend on the route of administration , which can be oral , topical , by inhalation or by injection , for example , intravenously . in one embodiment , the pharmaceutical composition is administered by intravenous infusion or injection . in another preferred embodiment , the pharmaceutical composition is administered by intramuscular or subcutaneous injection . the pharmaceutical composition for oral administration may be in the form of tablet , capsule , powder or liquid , for example , comprising an inert diluent or an assimilable edible carrier . a tablet can comprise a solid carrier , for example , gelatin or an adjuvant . a liquid pharmaceutical composition typically comprises a liquid carrier , for example , water , petroleum , animal or vegetable oil , mineral oil or synthetic oil . it is possible to include physiological saline solution , glucose or other sugar solutions , or glycols such as ethylene glycol , propylene glycol or polyethylene glycol . a specifically binding member ( and , when desired , other components ) may also be encapsulated into hard or soft shell gelatin capsules , compressed into tablets , or incorporated directly into the diet of a subject . for oral therapeutic administration , the active ingredient may be blended with excipients , and can be used in the form of ingestible tablets , buccal tablets , troches , capsules , elixirs , suspensions , syrups , wafer sorthe like . in order to administer the product of the present disclosure in routes other than parenteral administration , it may be necessary to coat a compound with a material that can prevent its deactivation or co - administer them . as to intravenous injection , or injection at a site of pain ( e . g ., tumor sites ), the active ingredient will be in the form of a parenterally acceptable aqueous solution , which is pyrogen - free and has a suitable pk , isotonicity and stability . a person skilled in the art can readily use , for example , an isotonic vehicle such as sodium chloride solution , ringer &# 39 ; s injection , or lactated ringer &# 39 ; s injection to prepare a suitable solution . when desired , preservatives , stabilizers , buffering agents , anti - oxidants and / or other additives can be contained . the agent or composition of the present disclosure can be used alone or in combination with an additional therapy simultaneously or sequentially , depending on the condition to be treated . the following examples are merely intended to illustrate the present disclosure in further detail and should by no means be construed as limiting the scope of the invention . a person skilled in the art will appreciate that modifications can be made to the following embodiments without departing from the scope , spirit and principle of the present disclosure as claimed by the appended claims . in the following examples , methods , apparatuses , reagents and protocols commonly used by those skilled in the art were employed , unless specified otherwise . b7 - h1 and pd - 1 were highly expressed in tumor microenvironment after radiation recurrence after rt is a common problem , which is at least partially due to the presence of rt - resistant tumor cells and / or tissues in a subject . the present inventor proposes that the course of recurrence , i . e ., progression of these rt - resistant tumors , may involve inhibitory molecules that inhibit t - cell responses . in order to investigate whether or not rt induces the expression of b7 - h1 / pd1 , the inventor stained b7 - h1 on tumor cells , dendritic cells and macrophages , and stained pd - 1 on cd4 30 t - cells and cd8 + t - cells . the experimental procedure was briefly described as follows . 5 × 10 5 tubo tumor cells ( derived from breast cancer cells of balb / c her2 / neu transgenic mice ) 18 were injected subcutaneously in the flanks of balb / c mice . at day 14 , the mice were locally irradiated with 15 grays ( gy ) using an x - ray generator ( pcm 1000 , pantak ). at day 28 , the tumors were removed and digested for 30 min with 0 . 2 mg / ml of collagenase to obtain a single - cell suspension for staining b7 - h1 with a standard staining protocol using 0 . 5 μg / ml of monoclonal antibody 10f . 9g2 purchased from bio - x cell , west lebanon , n . h . 03784 , usa . the inventor discovered that b7 - h1 was expressed not only on the tumor cells but also on dendritic cells and macrophages after radiation ( fig1 a and 1b ). the inventor also discovered that pd - 1 was highly expressed on infiltrative cd8 + t - cells and cd4 + t - cells ( fig1 c ). in order to investigate whether or not rt can induce expression of b7 - h1 on tumor cells ( myc - cap prostate cancer cell line 19 ), the tumor cells ( myc - cap tumor cell line ) were treated with 0 , 4 and 8 gy of irradiation . then , the irradiated tumor cells were cultured for 24 or 48 hours . after 24 or 48 hours , the cells were harvested , and the harvested cells ( 10 6 cells ) were subjected to standard staining with an anti - b7 - h1 monoclonal antibody ( 0 . 5 μg / ml of antibody 10f . 9g2 , purchased from bio - x cell , west lebanon , n . h . 03784 , usa ). the unirradiated myc - cap cells were used as a control . the results showed that rt significantly upregulated the expression of b7 - h1 in myc - cap cells . blockade of anti - b7 - h1 promoted local rt effect and reduced tumor burden in order to test whether or not rt - mediated b7 - h1 impairs acquired immune response , b7 - h1 / pd1 signaling pathway was blocked while performing rt . 5 × 10 5 tubo cells were injected subcutaneously in the flanks of balb / c mice . at day 14 , the mice were locally treated with a dosage of 12 gy of irradiation ( using an x - ray generator , pcm 1000 , pantak ). at days 15 , 18 and 21 , the mice were intraperitoneally injected with 50 μg of b7 - h1 blocking monoclonal antibody ( clone 10f . 9g2 , purchased from bio - x cell , west lebanon , n . h . 03784 , usa ), respectively , and tumor growth was monitored . the results showed that although neither rt nor the b7 - h1 blocking monoclonal antibody alone had any significant effect on tumor growth , the combination of rt and the antibody generated a synergistic effect , effectively causing significant tumor regression ( fig3 ). 1 . lee , y ., et al ., therapeutic effects of ablative radiation on local tumor require cd8 + t cells : changing strategies for cancer treatment . blood 114 , 589 - 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