WO2017202949A1 - Methods and compositions for treating cancers - Google Patents

Methods and compositions for treating cancers Download PDF

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Publication number
WO2017202949A1
WO2017202949A1 PCT/EP2017/062604 EP2017062604W WO2017202949A1 WO 2017202949 A1 WO2017202949 A1 WO 2017202949A1 EP 2017062604 W EP2017062604 W EP 2017062604W WO 2017202949 A1 WO2017202949 A1 WO 2017202949A1
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Prior art keywords
cells
cancer
pluripotent
population
cell
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PCT/EP2017/062604
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English (en)
French (fr)
Inventor
Frank Griscelli
Ali Turhan
Annelise BENNACEUR-GRISCELLI
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Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Descartes
Universite Paris Sud
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Assistance Publique Hopitaux de Paris APHP
Institut National de la Sante et de la Recherche Medicale INSERM
Universite Paris Descartes
Universite Paris Sud
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Priority to CN201780045481.XA priority Critical patent/CN109689089A/zh
Priority to RU2018141200A priority patent/RU2741786C2/ru
Priority to AU2017270234A priority patent/AU2017270234B2/en
Priority to CA3025057A priority patent/CA3025057A1/en
Priority to KR1020247020115A priority patent/KR20240103039A/ko
Priority to IL263224A priority patent/IL263224B2/en
Priority to BR112018074192A priority patent/BR112018074192A8/pt
Priority to US16/303,705 priority patent/US11458194B2/en
Priority to JP2018561512A priority patent/JP7563872B2/ja
Priority to CN202211701185.XA priority patent/CN116376812A/zh
Priority to KR1020187037459A priority patent/KR20190032295A/ko
Application filed by Assistance Publique Hopitaux de Paris APHP, Institut National de la Sante et de la Recherche Medicale INSERM, Universite Paris Descartes, Universite Paris Sud filed Critical Assistance Publique Hopitaux de Paris APHP
Priority to KR1020227044304A priority patent/KR102677451B1/ko
Priority to EP17726274.8A priority patent/EP3463433A1/en
Publication of WO2017202949A1 publication Critical patent/WO2017202949A1/en
Anticipated expiration legal-status Critical
Priority to JP2022110028A priority patent/JP7622006B2/ja
Priority to US17/815,586 priority patent/US20220370582A1/en
Priority to IL305882A priority patent/IL305882B2/en
Priority to AU2024200549A priority patent/AU2024200549A1/en
Priority to JP2024168499A priority patent/JP2025000812A/ja
Ceased legal-status Critical Current

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    • C12N5/0602—Vertebrate cells
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    • A61K31/19—Carboxylic acids, e.g. valproic acid
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Definitions

  • Embryonic stem cell-like gene expression and under-expression of Polycomb-regulated genes defining human pluripotent stem cell (ESC/IPSC) identity are associated in poorly differentiated human tumors with poor clinical outcome and distant recurrence after chemo-radiotherapy whatever the origin of cancers (breast, pancreas, bladder, lung, prostate, medulloblastoma ..). Poorly differentiated tumors with a "sternness" profile are related to mesenchymal traits on carcinoma cells with epithelial- mesenchymal "EMT" markers, low levels of MHC-I expression, immunosuppressive tumor microenvironment with pro-tumoral inflammatory leukocytes, stromal cells and macrophages.
  • EMT epithelial- mesenchymal
  • CSCs act as a reservoir to seed and replenish the tumor compartment. They also expand by self-renewal, disseminating to different tissues, generating metastases. These CSCs share pluripotent embryonic gene signatures and are resistant to anti-cancer drugs and radiotherapy. They also escape immune anti-tumor defenses for the reasons indicated above (immune-depressive micro-environment) .
  • Cancer- associated epigenetic aberrations are a characteristic trait of cancer stem cells involving every component of epigenetic machinery (DNA methylation, histone modifications, non-coding RNAs, specifically microRNA expression).
  • the invention thus relates to the use of a HDACi as an adjuvant, in particular for increasing the immune response against a vaccine composition, as well as to HDACi for its use as an adjuvant, or for increasing the immune response against a vaccine composition.
  • the invention also relates to the use of an HDACi for the manufacture of a vaccine composition containing one or more antigen(s) of interest, intended to have the patient develop an immune response against the antigen(s) of interest.
  • treating refers to both prophylactic or preventive treatment as well as curative or disease modifying treatment, including treatment of subject at high predisposed risk of contracting cancer such as hereditary family cancer syndromes or suspected to have contracted a cancer as well as subject who are ill or have been diagnosed as suffering from a cancer or medical condition, and includes suppression of clinical relapse.
  • the treatment may be administered to a subject having a cancer or who ultimately may acquire the cancer, in order to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of cancer or recurring cancer, or in order to prolong the survival of a subject beyond that expected in the absence of such treatment.
  • somatic cells are reprogrammed to induced pluripotent stem cells (iPSCs) by ectopic expression of defined factors such as Oct4, Sox2, Klf4 and c-My, or Oct4, Sox2, Lin28 and Nanog.
  • iPSCs induced pluripotent stem cells
  • the induced pluripotent stem cells are derived from mammals in particular (but not limited to) rodents, pigs, cats, dogs, and non-human primates, and human.
  • iPSCs have been successfully generated from somatic cells of various origins (fibroblast, blood cells, keratinoctytes%) and by using variable technologies (such as integrative lentivirus/ retrovirus and non integrative vectors such as sendai of virus, episomal vectors, synthetic mRNA, Adenovirus, rAAV, recombinant proteins%) with or without small chemical compounds.
  • variable technologies such as integrative lentivirus/ retrovirus and non integrative vectors such as sendai of virus, episomal vectors, synthetic mRNA, Adenovirus, rAAV, recombinant proteins.
  • the stem cells can be derived from mammals but not limited to rodents, pigs, cats, dogs, and primates, including humans.
  • the cell envelope integrity is maintained in step ii).
  • the pluripotent cells that are used in the method of treatment herein disclosed are inactivated.
  • the term "inactivated”, and grammatical variants thereof, is used herein to refer to a cell (e.g., a pluripotent cell) that is alive but has been rendered incapable of proliferation (i.e., mitotically inactivated).
  • the skilled in the art may use techniques that are known in the art including, but not limited to exposure to chemical agents, irradiation and/or lyophilization.
  • Pluripotent cells can be inactivated such that upon administration to a subject the pluripotent cells are incapable of dividing and thus cannot form teratomas in the subject.
  • cells present in a population of pluripotent cells are generally very homogenous when considering their genetic content (i.e. more than 95 % of the cells of the population present the same genetic background.
  • the inventors have shown that it is possible to design culture conditions that make it possible to induce DNA replication errors in pluripotent stem cells without triggering DNA damage- dependent apoptosis.
  • the pluripotent cells are very stable, application of the mutagen may have to be performed for a long period of time.
  • ENU when ENU is used, it may be applied for at least 7 days, more preferably at least 15 days, more preferably at least 20 days, more preferably at least 30 days, more preferably at least 40 days, more preferably at least 50 days or even at least 60 days.
  • the cells are washed (if the mutagen is a chemical agent) and can be further expanded, in the presence of the agent that favors MHC-I expression, in particular a HDACi. This agent is preferably also present during application of the mutagenic agent.
  • the vectors useful in the invention include, but are not limited to, naked plasmids, non-viral delivery systems (electroporation, sonoporation, cationic transfection agents, liposomes, etc .), phagemids, viruses, other vehicles derived from viral or bacterial sources that have been manipulated by the insertion or incorporation of the nucleic acid sequences.
  • Plasmid vectors have been extensively described in the art and are well known to those of skill in the art. See e.g. Sambrook et al., 1989. In the last few years, plasmid vectors have been used as DNA vaccines for delivering antigen-encoding genes to cells in vivo. They are particularly advantageous for this because they do not have the same safety concerns as with many of the viral vectors. These plasmids, however, having a promoter compatible with the host cell, can express a peptide from a gene operatively encoded within the plasmid.
  • homologous recombination refers to a gene targeting means for artificially modifying a specific gene on a chromosome or a genome.
  • genomic fragment having a portion homologous to that of a target sequence on the chromosome is introduced into cells, the term refers to recombination that takes place based on the nucleotide sequence homology between the introduced genomic fragment and the locus corresponding thereto on the chromosome.
  • This mutation rate of the genes is studied in the cell population, after exposure to the mutagenic agent, before or after further expansion, if such further expansion is performed.
  • Predisposition genes are, for instance (see also Lindor et al, 2008 Journal of the National Cancer Institute Monographs, No. 38, Concise Handbook of Familial Cancer Susceptibility Syndromes, Second Edition):
  • the histone deacetylase inhibitor is Chidamide (CS055/HB 1-8000) having the following chemical formula (C22H19FN4O2).
  • a “therapeutically effective amount” is intended for a minimal amount of active agent which is necessary to impart therapeutic benefit to a subject.
  • a “therapeutically effective amount” to a subject is such an amount which induces, ameliorates or otherwise causes an improvement in the pathological symptoms, disease progression or physiological conditions associated with or resistance to succumbing to a disorder. It will be understood that the total daily usage of the compounds of the present invention will be decided by the attending physician within the scope of sound medical judgment.
  • the specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; activity of the specific compound employed; the specific composition employed, the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific compound employed; the duration of the treatment; drugs used in combination or coincidental with the specific compound employed; and like factors well known in the medical arts.
  • the daily dosage of the products may be varied over a wide range from 0.01 to 1,000 mg per adult per day.
  • the subject is administered with i) a population of pluripotent cells and ii) a compound selected from a group which activates MHC expression and/or immune response, as a combined preparation and a chemotherapeutic agent.
  • chemotherapeutic agent refers to chemical compounds that are effective in inhibiting tumor growth.
  • examples of chemotherapeutic agents include alkylating agents such as thiotepa and cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethylenethiophosphaorarnide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a carnptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins
  • the immunotherapeutic treatment consists of an adoptive immunotherapy, as described by Nicholas P. Restifo, Mark E. Dudley and Steven A. Rosenberg ("Adoptive immunotherapy for cancer: harnessing the T cell response, Nature Reviews Immunology, Volume 12, April 2012).
  • adoptive immunotherapy the patient's circulating lymphocytes, or tumor- infiltrated lymphocytes, are isolated in vitro, activated by lymphokines such as IL-2 and readministered (Rosenberg et al., 1988; 1989).
  • the activated lymphocytes are most preferably be the patient's own cells that were earlier isolated from a blood sample and activated (or "expanded") in vitro.
  • the carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetables oils.
  • the proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants.
  • the prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like.
  • EXAMPLE 2 -Valproic acid modulates the expression of MHC class 1 and the expression of embryonic genes
  • MHC The major histocompatibility complex
  • the main function of MHC molecules is to bind to new and foreign antigens and to display them on the cell surface for recognition by the appropriate T- cells:
  • MHC class II By interacting with CD4 molecules on surfaces of helper T cells, MHC class II mediates establishment of specific immunity called acquired immunity or adaptive immunity.
  • CD8 molecules on surfaces of cytotoxic T cells By interacting with CD8 molecules on surfaces of cytotoxic T cells, MHC class I mediates destruction of infected or malignant host cells.
  • Immune tolerance is an important means by which growing tumors, which have mutated proteins and altered antigen expression, prevent elimination by the host immune system. Tumor immune tolerance can be explain in part by the absent of ⁇ 2- ⁇ on the cell surface and or the absence of MHC class I on tumor cell. It was shown that VPA is able to increase the expression of MHC class I on 4T1 cells at dose between 0.2 mM to 2 mM.
  • ENU Ethyl-N-Nitrosurea

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PCT/EP2017/062604 2016-05-25 2017-05-24 Methods and compositions for treating cancers Ceased WO2017202949A1 (en)

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