Patent Abstract:
an improved autovaccination method designed to prevent or treat various neoplastic diseases by inducing a systemic immune response against a tumor and its remote metastases , consisting of the induction of an immunogenic cell death in one or more targeted tumor sites with local radiation therapy , cryotherapy , heat , chemotherapy or various other treatments , followed by intratumoral / peritumoral injection of dsrnas in the same tumor site .

Detailed Description:
disclosed here is a method of treatment of human or veterinary neoplastic disease selected from among the group consisting of malignant brain tumors , melanoma , breast and lung cancer , colon cancer , sarcomas , renal cell cancer , leukemias , lymphomas , and other neoplasms , comprising the steps of : 1 ) administering to a primary or metastatic tumor site a suitable local tumoricidal agent such as radiation , chemotherapy , cryotherapy , embolization , or other agents resulting in release of tumor associated antigens , and 2 ) administering dsrna intratumorally or peritunorally in two or more doses spaced 4 - 72 hours apart , where the dose is in a moderate range ( preferably 5 to 100 mcg / kg in humans ) sufficient to induce measurable but not excessive levels of serum interferon and certain other cytokines and chemokines ; and for unblocking and stimulation of certain interferon and dsrna inducible enzyme systems , toll like receptor 3 , dendritic cells , antigen specific cytotoxic lymphocytes , memory t cells and other immune cells . human or veterinary diseases having a tumor site may be treated , according to the disclosed invention by administering to a primary or metastatic tumor site a suitable local tumoricidal agent and administering a double stranded rna molecule intratumorally or peritumorally . the double stranded rna molecule is preferably selected from the group consisting of poly - iclc , poly - ic ( poly inosinic - polycytidilic acid ), poly - au ( poly adenylic - poly uridylic acid ), or dsrna molecules with base modifications or modifications to the nucleic acid backbone , sugar moiety , or other sites in one or both strands of the nucleic acids , or which are incorporated in liposomes or polymers , and which bind to and / or activate immune cells through an interaction with the double stranded rna pattern recognition receptors ( prr ), including but not limited to toll - like receptor 3 . diseases which should respond to the treatment include malignant brain tumors , melanoma , breast and lung cancer , colon cancer , sarcomas , renal cell cancer , hepatoma , lymphomas , and other solid neoplasms . suitable tumoricidal agents include ionizing radiation , chemotherapy , cryotherapy , heat , embolization , or other agents resulting in release of tumor associated antigens . the invention will be described using the example of the double stranded rna molecule poly - iclc . while dosages , dose cycles and number of repetitions are determined by the specific patient according to methods known to those of skill in the art , it is expected that a suitable treatment would include intratumoral poly - iclc administered in a dose cycle comprising two or more doses spaced 4 - 72 hours apart , where the dose is in a moderate range ( 5 to 100 mcg / kg in humans ) sufficient to induce measurable but not excessive levels of serum interferon and certain other cytokines and chemokines ; and for unblocking and stimulation of certain interferon and dsrna inducible enzyme systems , toll like receptor 3 , dendritic cells , antigen specific cytotoxic lymphocytes , memory t cells and other immune cells . the dose cycles might be repeated weekly or twice weekly for up to one year . an alternate regime might be chemotherapy administered for 1 day to 3 weeks prior to intratumoral poly - iclc injection in order to decrease t regulatory cells and enhance the tumor - specific immune response . one suitable chemotherapeutic agent is cyclophosphamide , although the choice of specific agents would be patient specific and selected by methods known to those of skill in the art . an objective is to administer in a dose which is sufficient to activate the component immune cell or cells that leads to a return of the immune response to a normal disinhibited state or to an activated state , including but not limited to dendritic cells , cytotoxic t - cells , t - cells , or b - cells ; typically , this would include a dose of from 1 to 100 micrograms per kilogram of body weight , with a preferred dose of from 5 to 50 micrograms per kilogram of body weight . whether the quality of the cellular immune response is restored may be determined by measures such as generation of polyfunctional t - cells capable of a balanced cytokine and gamma interferon secretion . as alternatives , poly - iclc may be dosed alone , prior to its subsequent dosing in combination with autovaccination , one to several times or in combination multiple times , with the dosing regimen encompassing from one week to several years , or poly - iclc may be dosed alone , prior to its subsequent dosing in combination with a vaccine , one to several times or in combination with a vaccine multiple times in from one to multiple cycles that span a dosing regimen that encompasses at least one month . poly - iclc may be administered two to three times per week for a period extending from two days to four weeks prior to presentation of antigen or vaccine . in addition , poly - iclc may be combined with other pathogen associated molecular patterns ( pamps ). in addition to stimulation of dendritic cells and tumor specific cytotoxic t cells ( ctl ), poly - iclc also induces interleukin - 15 and other cytokines and chemokines that are important in the maintenance of specific ctl and immune memory cells , as well as in targeting of the remote tumor metastases . a closely related goal of the therapy is to improve the quality as well as the quantity of the immune response , including the generation of polyfunctional t - cells capable of a balanced cytokine and gamma interferon secretion . ( pantaleo and koup 2004 ) it is thus advantageous to follow the initial induction with subsequent poly - iclc treatments twice weekly for several weeks . in addition , repeated boosting ‘ autovaccination ’ cycles as described above every 4 to 8 weeks may be necessary to maintain tumor immunity . preconditioning with certain chemotherapeutic agents , including temozolomide and cyclophosphamide can paradoxically enhance the immune response to the autovaccination therapy . finally , this method could be extended by using other double - stranded rna molecules , including but not limited to poly - iclc , poly - ic , poly - au , dsrna molecules with base modifications or modifications to the nucleic acid backbone , sugar moiety , or other sites in one or both strands of the nucleic acids , or encased in liposomes or various polymers , and which bind to and / or activate immune cells through an interaction with the double stranded rna pattern recognition receptors ( prr ), including but not limited to toll - like receptor 3 . combination of poly - iclc with other pathogen associated molecular patterns ( pamps ) that activate other toll - like receptors has also been reported to boost the response to certain tumors or pathogens . clinical protocol : intratumoral poly - iclc plus cryotherapy in metastatic melanoma : a phase i / ii study patients with advanced metastatic melanoma with a karnofsdy performance score of 60 or better ( able to live at home and care for most personal needs ) will be selected for inclusion . a single superficial metastatic lesion will be identified for targeting . patients will first receive a single treatment with 2 - 3 mg / kg of cyclophosphamide preconditioning on day 1 . on day 2 the one targeted lesion will be frozen with liquid nitrogen in order to enhance tumor antigen release . this will be followed on days 2 and 4 with intratumoral / peritumoral injection of poly - iclc ( 0 . 5 to 1 mg ) additional poly - iclc will then be dosed , peritumorally in paired treatments two days apart on weeks 2 , 3 , and 4 . this will give a total of 8 poly - iclc treatments per cycle . at the end of week four ( and of week 9 and 14 ) there will be a one - week rest period during which patients will not receive any study medication . each additional cycle will thus begin after a one - week rest period , and the three treatment cycles are expected to take a total of 15 weeks . depending on patient tolerance and tumor response , additional cycles may be offered to the patient . primary outcome measures will be tumor response in remote metastases , and survival . clinical protocol : autologous vaccination against hepatic cancers with radiotherapy and intratumoral poly - iclc hepatocellular carcinomas ( hcc ) produce factors blocking the host immune response and promoting immunologic tolerance to tumor antigens . intratumoral poly - iclc enhances dendritic cell maturation and tumor antigen uptake , generation of specific cytotoxic lymphocytes , and targeting of tumor cells through alteration of the local tumor microenvironment , including various chemokines . we report preliminary data from an ongoing phase i / ii clinical trial ( de la torre , salazar , et al .) using : 1 ) low dose 3d conformal radiation ( 3drt ) to increase hcc tumor antigen release , 2 ) ultrasound guided intra / peri tumor injection of poly - iclc , ( poly - iclc ) to initiate an innate and adaptive immune response in the local tumor environment , 3 ) percutaneous arterial embolization of the targeted lesion , and 4 ) systemic im poly - iclc to enhance immunologic memory and targeting day 1 , 2 , 3 : 2 . 5 gray 3drt . day 4 : ultrasound guided peri / intra - tumoral poly - iclc ( 0 . 25 to 1 mg in 5 cc saline ) & amp ; embolization ; twice weekly for three weeks : intramuscular poly - iclc ( 20 mcg / kg ) patients have tolerated the treatment without significant side effects and remain clinically well . currently , no new lesions have been detected on follow - up triple phase ct in the 4 patients completing the protocol . two patients have shown radiologic response in remote lesions that were not originally targeted by radiation , indicating a systemic immune response that is capable of targeting remote metastases . these early data show that tumor conditioning with radiation , intra - tumoral injection of poly - iclc tlr3 ligand into liver cancers , followed by tumor arterial embolization is feasible and safe in humans . stabilization and / or response in these typically very aggressive tumors is particularly promising and further supports the concept behind this disclosure . brody j , a . w ., cerwinski d , advani r , horning s j , ganjoo r , levy r ( 2008 ). “ clinical and immonologic responses to a novel in situ lymphoma vaccine maneuver : preliminary results of a phase ii trial of intratumoral cpg 7909 .” j clin oncology 26 ( may 20 suppl , abstr 3003 ). furumoto , k ., l . soares , et al . 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