Combination therapy

Abstract

Provided herein are combination therapies comprising either an epoxytigliane compound (e.g., tigilanol tiglate) and radiation, or a chemotherapeutic agent that is either i) a chemotherapeutic agent that damages DNA (e.g., cisplatin), or ii) a chemotherapeutic agent that inhibits tumor-associated host-derived cells that support tumor cell proliferation and / or invasion (e.g., 5-fluorouracil, or doxorubicin). Pharmaceutical compositions and kits comprising the epoxytigliane compound and a chemotherapeutic agent are also provided.

Description

[Technical field]

[0001] This application claims priority to Australian Provisional Patent Application No. 2022900340, filed on February 17, 2022, the contents of which are incorporated herein by reference in their entirety.

[0002] Field The present invention relates to combination therapies comprising epoxytigliane compounds and cancer chemotherapeutic agents or irradiation. Pharmaceutical compositions and kits comprising epoxytigliane compounds and cancer chemotherapeutic agents are also described. [Background technology]

[0003] background Treatment of cancer includes many options, including chemotherapy agents, radiation therapy, surgery, and immune stimulants. Some treatments are developed for specific types of cancer or cancers characterized by specific biological markers. Known treatments frequently have significant side effects and may not be suitable for some patients. For example, some tumors are inoperable, and some patients may have comorbidities that reduce the effectiveness of chemotherapy agents or prevent the use of certain therapeutic agents due to side effects.

[0004] One option to reduce side effects, expand the range of treatments available to patients with comorbidities, and reduce the development of resistance to certain treatments is to use combinations of therapeutic agents. The effect of combinations of therapeutic agents may be additive, synergistic, or antagonistic, and it is not always possible to predict whether a combination may be additive, synergistic, or antagonistic, or whether a reduction in the therapeutic dose would be effective.

[0005] There is a need for combination therapies that provide patient benefit, such as reduced dosages, fewer side effects, synergistic effects, increased treatment options, and / or improved rates of treatment success.

[0006] Epoxytigliane compounds have potent antitumor activity. When administered intratumorally, epoxytiglianes induce rapid hemorrhagic necrosis of the tumor mass by directly destroying tumor vasculature (Boyle et al. 2014).

[0007] The present disclosure is based, at least in part, on the discovery that compound 1, an epoxytigliane compound, can provide a synergistic effect with cisplatin, 5-fluorouracil, doxorubicin and radiation in a variety of cancers, including squamous cell carcinoma, adenocarcinoma and fibrosarcoma. Summary of the Invention

[0008] overview In a first aspect, a method of treating a tumor is provided comprising administering a combination of an epoxytigliane compound and a second therapeutic agent; the epoxytigliane compound is a compound of formula (I): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 is alkyl; R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl; R6 is hydrogen or -R 10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 is alkyl; R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10 Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R 11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10 AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11 , -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] and; R 10-C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) chemotherapy agents that damage DNA; ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; and iii) irradiation, is selected from.

[0009] In another embodiment, a combination of an epoxytigliane compound and a second therapeutic agent for use in treating a tumor is provided; the epoxytigliane compound is a compound of formula (I): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 is alkyl; R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl; R6 is hydrogen or -R10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 is alkyl; R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20 Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10 Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R 11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11 , -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] and; R 10 -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) chemotherapy agents that damage DNA; ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; and iii) irradiation; is selected from.

[0010] In a further aspect, there is provided the use of an epoxytigliane compound in the manufacture of a first medicament, and a second chemotherapeutic agent in the manufacture of a second medicament, as a combination therapy for treating a tumor; said epoxytigliane compound being a compound of formula (I): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 is alkyl; R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl; R6 is hydrogen or -R 10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 is alkyl; R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20 Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10 Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R 11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10 AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11 , -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] and; R 10 -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) DNA damaging chemotherapeutic agents; and ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; is selected from.

[0011] In yet another embodiment, there is provided the use of an epoxytigliane compound in the manufacture of a medicament in combination therapy with radiation for treating a tumor; said epoxytigliane compound being a compound of formula (I): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 is alkyl; R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl; R6 is hydrogen or -R 10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 is alkyl; R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20 Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10 Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10 AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11 , -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] and; R 10 -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted. [Brief description of the drawings]

[0012] [Figure 1a] FIG. 1a is a plot of individual SCC-15 tumor volumes over time following combined treatment with intratumoral Compound 1, intraperitoneal cisplatin and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Figure 1b] FIG. 1b is a plot of mean SCC-15 tumor volume over time following combined treatment with intratumoral Compound 1, intraperitoneal cisplatin and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Diagram 2] FIG. 2 is a plot of Kaplan-Meier survival curves of SCC-15 tumors over time following combined treatment with intratumoral Compound 1, intraperitoneal cisplatin and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Figure 3a] FIG. 3a is a plot of individual UV-13-1 tumor volumes over time following combined treatment with intratumoral Compound 1, intraperitoneal cisplatin and control agents in a C3H / HeNCr syngeneic mouse model. [Figure 3b] FIG. 3b is a plot of mean UV-13-1 tumor volume over time following combined administration of intratumoral Compound 1, intraperitoneal cisplatin and control agents in a C3H / HeNCr syngeneic mouse model. [Figure 4] FIG. 4 is a plot of Kaplan-Meier survival curves of UV-13-1 tumors over time following combined administration of intratumoral Compound 1, intraperitoneal cisplatin and control agents in a C3H / HeNCr syngeneic mouse model. [Figure 5a]FIG. 5a is a plot of individual SCC-15 tumor volumes over time following combined treatment with intratumoral Compound 1, external beam radiation, and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Figure 5b] FIG. 5b is a plot of mean SCC-15 tumor volume over time following combined treatment with intratumoral Compound 1, external beam radiation, and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Figure 6] FIG. 6 is a plot of Kaplan-Meier survival curves of SCC-15 tumors over time following combined treatment with intratumoral Compound 1, external beam radiation, and control agents in a BALB / c Foxn1nu xenogeneic mouse model. [Figure 7a] FIG. 7a is a plot of individual MC38 tumor volumes over time following combined treatment with intratumoral Compound 1, external beam radiation, and control agents in a C57BL / 6 syngeneic mouse model. [Figure 7b] FIG. 7b is a plot of mean MC38 tumor volume over time following combined treatment with intratumoral Compound 1, external beam radiation, and control agents in a C57BL / 6 syngeneic mouse model. [Figure 8] FIG. 8 is a plot of Kaplan-Meier survival curves of MC38 tumors over time following combined treatment with intratumoral Compound 1, intraperitoneal cisplatin and control agents in a C57BL / 6 syngeneic mouse model. [Figure 9a] FIG. 9a plots cell viability of A431 cells following combination treatment with cisplatin and different concentrations of Compound 1, with each data point representing the average of three different experiments. [Figure 9b] FIG. 9b plots cell viability of SCC-15 cells following combination treatment with cisplatin and different concentrations of Compound 1, with each data point representing the average of three different experiments. [Figure 10a] FIG. 10a plots cell viability of A431 cells following combination treatment with 5-fluorouracil and different concentrations of Compound 1, with each data point representing the average of three different experiments. [Figure 10b] FIG. 10b plots cell viability of SCC-15 cells following combination treatment with 5-fluorouracil and different concentrations of Compound 1, with each data point representing the average of three different experiments. [Figure 11a] FIG. 11a plots cell survival of A431 cells after combined treatment with Compound 1 and different doses of radiation, with each data point representing the average of three different experiments. [Figure 11b] FIG. 11b plots cell survival of SCC-15 cells after combined treatment with Compound 1 and different doses of radiation, with each data point representing the average of three different experiments. [Figure 12] FIG. 12 is a plot of the combination effect (Fa-CI) of Compound 1 and cisplatin, 5-fluorouracil, and radiation in A431 cells. [Figure 13] FIG. 13 is a plot of the combination effect (Fa-CI) of Compound 1 and cisplatin, 5-fluorouracil, and radiation in SCC-15 cells. [Figure 14a] FIG. 14a is a plot of Kaplan-Meier survival curves over time for B16-F10-OVA tumors with tumor size greater than 100 mm3 after combined treatment with intratumoral compound 1 (5 μg or 10 μg), intraperitoneal doxorubicin and control drug in C57BL / 6 mice. [Figure 14b] FIG. 14b is a plot of Kaplan-Meier survival curves over time for B16-F10-OVA tumors (overall survival) following combined treatment with intratumoral Compound 1 (5 μg or 10 μg), intraperitoneal doxorubicin and control agents in C57BL / 6 mice. [Figure 15a] FIG. 15a is a plot of Kaplan-Meier survival curves over time for Lewis Lung Carcinoma (LLC) tumors with tumor size greater than 100 mm3 after combined treatment with intratumoral Compound 1 (5 μg or 10 μg), intraperitoneal doxorubicin and control agents in C57BL / 6 mice. [Figure 15b]FIG. 15b is a plot of Kaplan-Meier survival curves over time for Lewis Lung Carcinoma (LLC) tumors (overall survival) following combined treatment with intratumoral Compound 1 (5 μg or 10 μg), intraperitoneal doxorubicin and control agents in C57BL / 6 mice. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] Detailed Description definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, preferred methods and materials are described. For purposes of the present invention, the following terms are defined below.

[0014] This disclosure refers to the entire contents of certain documents incorporated herein by reference. In the event of a conflict between the teachings of this disclosure and the contents of those documents, the teachings of this disclosure shall control.

[0015] Where a prior art document is referred to in this specification, it should be understood that such reference does not constitute an admission that the document forms part of the general knowledge in the art in Australia or anywhere else.

[0016] As used herein, the articles "a" and "an" are used to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, "an element" means one element or more than one element.

[0017] As used herein, the term "about" refers to a quantity, level, value, dimension, size, or amount that varies by as much as 25%, 20%, 15%, or 10% relative to a reference quantity, level, value, dimension, size, or amount.

[0018] As used herein, the term "and / or," e.g., "X and / or Y," is to be construed as meaning either "X and Y" or "X or Y," and is to be considered as clearly endorsing both meanings or either meaning.

[0019] Throughout this specification, unless the context requires otherwise, the words "comprise", "comprises" and "comprising" shall be construed as meaning the inclusion of certain steps or elements or groups of steps or elements and not as meaning the exclusion of other steps or elements or groups of steps or elements.

[0020] Unless otherwise indicated, the terms "first," "second," etc. are used herein merely as labels, and are not intended to impose any sequential, positional, or hierarchical requirements on the items to which they refer. Moreover, a reference to a "second" item does not require or preclude the presence of a lower-numbered item (e.g., a "first" item) and / or a higher-numbered item (e.g., a "third" item).

[0021] As used herein, the phrase "at least one" when used in conjunction with a list of items means that one or more different combinations of the items in the list may be used, and only one of the items in the list may be required. The items may be specific objects, things, or categories. In other words, "at least one" means that any combination or number of items in the list may be used, but not all items in the list are required. For example, "at least one of item A, item B, and item C" may mean item A; item A and item B; item B; item A, item B, and item C; or item B and item C. In some cases, "at least one of item A, item B, and item C" may mean, for example, but not limited to, two items A, one item B, and ten items C; four items B and seven items C; or some other suitable combination.

[0022] The term "alkyl" refers to optionally substituted straight and branched chain hydrocarbons having 1 to 20 carbon atoms. Where appropriate, the alkyl group can have a specific number of carbon atoms, such as C1-C6 alkyl, including alkyl groups having 1, 2, 3, 4, 5, or 6 carbon atoms in a straight or branched arrangement. Non-limiting examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, s- and t-butyl, pentyl, 2-methylbutyl, 3-methylbutyl, hexyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-ethylbutyl, 3-ethylbutyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, and pentadecyl.

[0023] The term "alkenyl" refers to an optionally substituted, unsaturated, straight or branched chain hydrocarbon having 2 to 20 carbon atoms and at least one double bond. Where appropriate, the alkenyl group can have a specific number of carbon atoms, such as C2-C6 alkenyl, including alkenyl groups having 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Non-limiting examples of alkenyl groups include ethenyl, propenyl, isopropenyl, butenyl, s- and t-butenyl, pentenyl, hexenyl, hepto-1,3-diene, hexa-1,3-diene, nona-1,3,5-triene, and the like.

[0024] The term "alkynyl" refers to an optionally substituted, unsaturated, straight or branched chain hydrocarbon having 2-20 carbon atoms and at least one triple bond. Where appropriate, the alkynyl group can have a specific number of carbon atoms, such as C2-C6 alkynyl, including alkynyl groups containing 2, 3, 4, 5 or 6 carbon atoms in a straight or branched arrangement. Non-limiting examples include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and the like.

[0025] The terms "cycloalkyl" and "carbocyclic" refer to an optionally substituted, saturated or unsaturated, monocyclic, bicyclic, or tricyclic hydrocarbon group. Where appropriate, cycloalkyl groups can have a specific number of carbon atoms, e.g., C3-C6 cycloalkyl is a carbocyclic group having 3, 4, 5, or 6 carbon atoms. Non-limiting examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like.

[0026] "Aryl" means a monocyclic, bicyclic or tricyclic carbocyclic ring system having up to seven atoms in each ring, with at least one ring being aromatic. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tetrahydronaphthyl, indanyl and biphenyl. An aryl may contain one to three benzene rings. When more than one aromatic ring is present, the rings may then be fused together such that adjacent rings share a common bond.

[0027] Each alkyl, alkenyl, alkynyl, cycloalkyl or aryl, whether as an individual component or as part of a larger component, may optionally be selected from the group consisting of C 1-6 Alkyl, C 2-6 Alkenyl, C 3-6 Cycloalkyl, oxo (=O), OH, -SH, C 1-6 Alkyl O-, C 2-6 Alkenyl O-, C 3-6 CycloalkylO-,C 1-6 Alkyl S-, C 2-6 Alkenyl S-, C 3-6 CycloalkylS-, -CO2H, -CO2C 1-6 Alkyl, -NH2, -NH(C 1-6 alkyl), -N(C 1-6 alkyl)2, -NH(phenyl), -N(phenyl)2, -CN, -NO2, -halogen, -CF3, -OCF3, -SCF3, -CHF2, -OCHF2, -SCHF2, -phenyl, -C 1-6 Alkylphenyl, -Ophenyl, -C(O)phenyl, -C(O)C 1-6Examples of suitable substituents include, but are not limited to, methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, vinyl, methoxy, ethoxy, propoxy, isopropoxy, butoxy, methylthio, ethylthio, propylthio, isopropylthio, butylthio, hydroxy, hydroxymethyl, hydroxyethyl, hydroxypropyl, hydroxybutyl, fluoro, chloro, bromo, iodo, cyano, nitro, -CO2H, -CO2CH3, -C(O)CH3, trifluoromethyl, trifluoromethoxy, trifluoromethylthio, difluoromethyl, difluoromethoxy, difluoromethylthio, morpholino, amino, methylamino, dimethylamino, phenyl, phenoxy, phenylcarbonyl, benzyl, and acetyl.

[0028] The epoxytigliane compounds may be in the form of pharma- ceutically acceptable salts, however, it will be understood that non-pharma- ceutically acceptable salts are also included within the scope of the present invention, as they may be useful as intermediates in the preparation of pharma- ceutical acceptable salts, or may be useful during storage and transportation. Suitable pharma- ceutically acceptable salts include, but are not limited to, salts of pharma- ceutically acceptable inorganic acids, such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, sulfamic acid, and hydrobromic acid, or salts of pharma- ceutically acceptable organic acids, such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, maleic acid, citric acid, lactic acid, muconic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edetic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid.

[0029] Base salts include, but are not limited to, those formed with pharma- ceutically acceptable cations, such as sodium, potassium, lithium, calcium, magnesium, ammonium, and alkylammonium.

[0030] Basic nitrogen-containing groups may be quaternized with agents such as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides and iodides; dialkyl sulfates, such as dimethyl sulfate and diethyl sulfate; and others.

[0031] It will also be appreciated that epoxytigliane compounds may have asymmetric carbons and therefore may exist in multiple stereoisomeric forms. Thus, the present invention also relates to substantially pure isomeric forms at one or more asymmetric carbons, such as those with an ee of greater than about 90%, such as those with an ee of about 95% or 97% or those with an ee of greater than 99%, as well as mixtures, including racemic mixtures. Such isomers may be obtained by isolation from natural sources, for example, asymmetric synthesis using chiral intermediates, or chiral resolution. The compounds of the present invention may exist as geometric isomers. The present invention also relates to compounds in substantially pure cis (Z) or trans (E) form or mixtures thereof.

[0032] The compounds of the invention may be obtained by isolation from plants or plant parts, derivatization of the isolated compounds, or derivatization of related compounds. Isolation and derivatization procedures are described in WO 2007 / 070985 and WO2014 / 169356.

[0033] The term "epoxytigliane compounds" generally refers to compounds having the following basic carbocyclic structure: [ka]

[0034] The compounds have a tricyclo[9.3.0.0]tetradecane system with a fused cyclopropane ring attached to a six-membered ring. An epoxide is fused to the seven-membered ring at the 6,7 positions. Epoxy tigliane compounds are generally epoxy tiglien-3-one compounds. The term "epoxy tiglien-3-one compounds" refers to compounds having the epoxy tigliane structure defined above, where the five-membered ring has the following 1,2-ene-3-one structure: [ka]

[0035] Throughout this specification, the term "epoxy tigliane compound" is used to refer to the epoxy tigliane compound of formula (I).

[0036] As used herein, the term "combination" refers to the administration of an epoxytigliane compound and a second chemotherapeutic agent (e.g., a cancer chemotherapeutic agent) in a single composition, or the administration of the second chemotherapeutic agent (e.g., a cancer chemotherapeutic agent) or radiation separately, simultaneously, or sequentially. The epoxytigliane compound and the second chemotherapeutic agent (e.g., a cancer chemotherapeutic agent) or radiation may be administered at different times and with different frequencies, but when administered in combination, they exert a biological effect at the same time or at overlapping times.

[0037] Treatment method The present invention relates to a method for treating tumors, the method comprising administering an epoxytigliane compound or a pharma- ceutically acceptable salt thereof in combination with a second chemotherapeutic agent or radiation.

[0038] In some embodiments, the tumor being treated is one in which the epoxytigliane compound can be delivered locally directly to the tumor. In certain embodiments, the tumor is a cutaneous tumor or a subcutaneous tumor or a tumor accessible from outside the body, e.g., a palpable tumor. In other embodiments, the tumor is an internal tumor. In some embodiments in which the tumor is an internally located tumor, localized delivery is achieved during surgery in which the tumor is exposed and the epoxytigliane compound can be injected. In other embodiments, the tumor is located internally and the epoxytigliane compound is delivered by injection guided by image-guided techniques, e.g., endoscopic ultrasound or stereotactic image guidance.

[0039] In some embodiments, the epoxytigliane compound is delivered systemically to one or more tumors.

[0040] In some embodiments, the tumor is a benign tumor. In other embodiments, the tumor is a malignant tumor. In some embodiments, the tumor is a primary tumor, and in other embodiments, the tumor is a metastatic tumor. In some embodiments, the tumor is a skin tumor. Examples of skin tumors include seborrheic keratosis, actinic keratosis, basal cell carcinoma (BCC), including nodular BCC, superficial BCC, invasive BCC, and micronodular BCC, squamous cell carcinoma (SCC), including in-situ SCC and invasive SCC, adenocarcinoma, melanoma, including superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma, acral lentigo melanoma, and desmoplastic / neutrophilic melanoma, sarcoma, cutaneous B-cell lymphoma, and cutaneous T-cell lymphoma. Examples of cutaneous and subcutaneous tumors that may be treated include hemangioma, telangiectatic granuloma, cherry hemangioma, glomus hemangioma, angiosarcoma, Karposi's sarcoma, Ewing's sarcoma, malignant fibrous histiocytoma, leiomyosarcoma, rhabdomyosarcoma, lipo-osteosarcoma, synovial sarcoma, stromal sarcoma, gastrointestinal stromal sarcoma, malignant peripheral nerve sheath tumor, primitive neuroectodermal tumor, neurofibroma, Merkel cell carcinoma, dermatofibroma, fibrosarcoma, epithelioid sarcoma, and mastocytoma (mast cell tumour).

[0041] When the tumor is a visceral tumor, the visceral tumor may be any tumor that is accessible by injection during surgery or by guided injection or that may be treated with systemic administration of epoxytigliane, and may include tumors of the brain, lung, colon, epidermoid, squamous, bladder, stomach, pancreas, breast, head, neck, renal system, kidney, liver, ovary, prostate, uterus, esophagus, testicle, cervix, vagina, thyroid or skin.

[0042] In some embodiments, the tumor is a head or neck tumor. In some embodiments, the head or neck tumor is a skin tumor. In other embodiments, the head or neck tumor is a mucosal epithelial tumor, e.g., arising in the mucosal epithelium of the upper aerodigestive tract. In certain embodiments, the tumor is head and neck squamous cell carcinoma (HNSCC).

[0043] In some embodiments, the tumor is a mucosal head or neck cancer, particularly a mucosal squamous cell carcinoma (SCC). In some embodiments, the mucosal tumor is associated with smoking and / or alcohol use. In some embodiments, the tumor is of the oropharynx, particularly of the tonsils or base of the tongue.

[0044] In some embodiments, the tumor is associated with human papilloma virus (HPV). In certain embodiments, the HPV-associated tumor is a mucosal SCC. HPV-associated mucosal SCC includes tumors of the oropharynx, vulva, vagina, and cervix.

[0045] In some embodiments, the tumor is a non-melanoma skin cancer (NMSC). NMSC can occur anywhere on the body. The tumor can be associated with UV exposure, skin phenotype, immunosuppression, xeroderma pigmentosum, chemical exposure, chronic ulcers and scars, ionizing radiation, or a combination thereof. In some embodiments, the tumor is a high-risk tumor having one or more of the following: size greater than 4 cm, depth >2 mm, presence of perineural or lymphatic invasion, involvement of the ear or lip, patient immunosuppression, and prior radiation therapy.

[0046] In some embodiments, the tumor is a squamous cell tumor (SCC), such as thyroid cancer, esophageal cancer, SCC of the lung, SCC of the penis (potentially associated with HPV), including Bowen's disease, erythroplasia of Queyrat and Bowenoid papulosis, prostate cancer, vaginal cancer, cervical cancer and bladder cancer, particularly bladder cancer associated with schistosomiasis.

[0047] In some embodiments, the tumor is a sarcoma, such as angiosarcoma, Karposi's sarcoma, Ewing's sarcoma, leiomyosarcoma, rhabdomyosarcoma, liposarcoma, synovial sarcoma, stromal sarcoma, gastrointestinal stromal sarcoma, fibrosarcoma, or epithelioid sarcoma. In some embodiments, the sarcoma is ultraviolet light-induced fibrosarcoma.

[0048] In other embodiments, the tumor is an adenocarcinoma. Examples of adenocarcinoma include esophageal cancer, pancreatic cancer, prostate cancer, cervical cancer, gastric cancer, invasive ductal carcinoma, ductal carcinoma in situ, invasive lobular carcinoma, colon cancer, colorectal cancer, adenocarcinoma of the lung, cholangiocarcinoma (bile duct cancer), and vaginal cancer.

[0049] Epoxytigliane Compounds The epoxytigliane compound is a 6,7-epoxytigli-1,2-en-3-one compound of formula (I): [ka] or an isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 is alkyl; R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl; R6 is hydrogen or -R 10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 is alkyl; R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20 Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10 Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R 11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10 AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11, -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] and; R 10 -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted.

[0050] In some embodiments, the epoxytigliane compound of formula (I) is a compound of formula (II): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof, wherein R6, R7, and R9 are as defined in formula (I).

[0051] In certain embodiments of formula (I) or (II), one or more of the following may be present: R1 is -C 1-3 Alkyl, in particular -CH3; R2 is -OC(O)C 1-20 Alkyl, -OC(O)C 2-20Alkenyl, -OC(O)C 2-20 Alkynyl, -OC(O)cycloalkyl, -OC(O)C 1-10 Alkylcycloalkyl; -OC(O)C 2-10 Alkenylcycloalkyl, -OC(O)C 2-10 Alkynylcycloalkyl, -OC(O)aryl, -OC(O)C 1-10 Alkylaryl, -OC(O)C 2-10 Alkenylaryl, -OC(O)C 2-10 Alkynylaryl, -OC(O)C 1-10 AlkylC(O)R 11 , -OC(O)C 2-10 AlkenylC(O)R 11 , -OC(O)C 2-10 AlkynylC(O)R 11 , -OC(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -OC(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -OC(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -OC(O)C 1-10 Alkyl SR 11 , -OC(O)C 2-10 Alkenyl SR 11 , -OC(O)C 2-10 Alkynyl SR 11 , -OC(O)C 1-10 AlkylC(O)OR 11 , -OC(O)C 2-10 AlkenylC(O)OR 11 , -OC(O)C 2-10 AlkynylC(O)OR 11 , -OC(O)C 1-10 AlkylC(O)SR 11 , -OC(O)C 2-10 AlkenylC(O)SR 11 , and -OC(O)C 2-10 AlkynylC(O)SR 11 ;In particular, -OC(O)C 1-20 Alkyl, -OC(O)C 2-20Alkenyl, -OC(O)C 2-20 Alkynyl, -OC(O)cycloalkyl, -OC(O)C 1-10 Alkylcycloalkyl; -OC(O)C 2-10 Alkenylcycloalkyl, -OC(O)C 2-10 Alkynylcycloalkyl or -OC(O)aryl; more particularly -OC(O)C 1-20 Alkyl, -OC(O)C 2-20 Alkenyl or -OC(O)C 2-20 Alkynyl; R3 is -OC(O)C 1-20 Alkyl, -OC(O)C 2-20 Alkenyl, -OC(O)C 2-20 Alkynyl, -OC(O)cycloalkyl, -OC(O)C 1-10 Alkylcycloalkyl; -OC(O)C 2-10 Alkenylcycloalkyl, -OC(O)C 2-10 Alkynylcycloalkyl, -OC(O)aryl, -OC(O)C 1-10 Alkylaryl, -OC(O)C 2-10 Alkenylaryl, -OC(O)C 2-10 Alkynylaryl, -OC(O)C 1-10 AlkylC(O)R 11 , -OC(O)C 2-10 AlkenylC(O)R 11 , -OC(O)C 2-10 AlkynylC(O)R 11 , -OC(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -OC(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -OC(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -OC(O)C 1-10 Alkyl SR 11 , -OC(O)C 2-10 Alkenyl SR 11 , -OC(O)C 2-10 Alkynyl SR 11 , -OC(O)C1-10 AlkylC(O)OR 11 , -OC(O)C 2-10 AlkenylC(O)OR 11 , -OC(O)C 2-10 AlkynylC(O)OR 11 , -OC(O)C 1-10 AlkylC(O)SR 11 , -OC(O)C 2-10 AlkenylC(O)SR 11 , and -OC(O)C 2-10 AlkynylC(O)SR 11 ;In particular, -OC(O)C 1-20 Alkyl, -OC(O)C 2-20 Alkenyl, -OC(O)C 2-20 Alkynyl, -OC(O)cycloalkyl, -OC(O)C 1-10 Alkylcycloalkyl; -OC(O)C 2-10 Alkenylcycloalkyl, -OC(O)C 2-10 Alkynylcycloalkyl or -OC(O)aryl; more particularly -OC(O)C 1-20 Alkyl, -OC(O)C 2-20 Alkenyl or -OC(O)C 2-20 Alkynyl; R4 and R5 are independently -C 1-3 alkyl, in particular, selected from -CH3; R6 is hydrogen, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl or -C(O)aryl; in particular hydrogen, -C(O)C 1-3 Alkyl, -C(O)C 2-3 Alkenyl or -C(O)C 2-3 alkynyl, more especially hydrogen or -C(O)CH3; R7 is hydroxyl, -OC(O)C 1-6 Alkyl, -OC(O)C 2-6 Alkenyl or -OC(O)C 2-6 Alkynyl; especially hydroxyl, -OC(O)C 1-3 Alkyl, -OC(O)C 2-3 Alkenyl or -OC(O)C2-3 alkynyl, more particularly hydroxyl or -OC(O)CH; and R8 is -C 1-3 alkyl, in particular -CH; applies.

[0052] In some embodiments, the epoxytigliane compound of formula (I) is a compound of formula (Ia): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen or C 1-6 alkyl (e.g., R1 is C 1-3 alkyl, e.g., CH3, etc.); R2 is -OH or -OR9; R3 is -OH or -OR9; R4 and R5 are independently hydrogen and C 1-6 alkyl (e.g., R and R are independently selected from C 1-3 alkyl, e.g., CH3, etc.); R6 is hydrogen or -R 10 and; R7 is hydroxy or OR 10 and; R8 is hydrogen or C 1-6 alkyl (e.g., R is C 1-3 alkyl, e.g., CH3, etc.); R9 is -C 1-20 Alkyl, -C 2-20 Alkenyl, -C 2-20 Alkynyl, -C(O)C 1-20 Alkyl, -C(O)C 2-20 Alkenyl, -C(O)C 2-20 Alkynyl, -C(O)cycloalkyl, -C(O)C 1-10 Alkylcycloalkyl; -C(O)C 2-10 Alkenylcycloalkyl, -C(O)C 2-10Alkynylcycloalkyl, -C(O)aryl, -C(O)C 1-10 Alkylaryl, -C(O)C 2-10 Alkenylaryl, -C(O)C 2-10 Alkynylaryl, -C(O)C 1-10 AlkylC(O)R 11 , -C(O)C 2-10 AlkenylC(O)R 11 , -C(O)C 2-10 AlkynylC(O)R 11 , -C(O)C 1-10 Alkyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 Alkenyl CH(OR 11 )(OR 11 ), -C(O)C 2-10 AlkynylCH(OR 11 )(OR 11 ), -C(O)C 1-10 Alkyl SR 11 , -C(O)C 2-10 Alkenyl SR 11 , -C(O)C 2-10 Alkynyl SR 11 , -C(O)C 1-10 AlkylC(O)OR 11 , -C(O)C 2-10 AlkenylC(O)OR 11 , -C(O)C 2-10 AlkynylC(O)OR 11 , -C(O)C 1-10 AlkylC(O)SR 11 , -C(O)C 2-10 AlkenylC(O)SR 11 , -C(O)C 2-10 AlkynylC(O)SR 11 , [ka] (For example, R9 is -C(O)C 1-20 Alkyl or -C(O)C 2-20 alkenyl, such as tigloyl, 2-methylbutanoyl, hexanoyl, myristoyl, propanoyl or acetyl; R10 -C 1-6 Alkyl, -C 2-6 Alkenyl, -C 2-6 Alkynyl, -C(O)C 1-6 Alkyl, -C(O)C 2-6 Alkenyl, -C(O)C 2-6 Alkynyl, -C(O)aryl, -C(O)C 1-6 Alkylaryl, -C(O)C 2-6 Alkenylaryl, or -C(O)C 2-6 alkynylaryl (e.g., R 10 is -C(O)C 1-3 alkyl, such as -C(O)CH3; and R 11 is hydrogen, -C 1-10 Alkyl, -C 2-10 Alkenyl, -C 2-10 It is alkynyl, cycloalkyl or aryl.

[0053] In some embodiments, the epoxytigliane compound of formula (I) is a compound of formula (Ib): [ka] or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is C 1-3 alkyl (e.g., R1 is CH3); R2 is -OR9; R3 is -OR9; R4 and R5 are independently C 1-3 alkyl; R6 is hydrogen or -R 10 and; R7 is hydroxy; R8 is C 1-3 is alkyl; R9 is -C(O)C 1-20 Alkyl or -C(O)C 2-20alkenyl (e.g., R9 is tigloyl, 2-methylbutanoyl, hexanoyl, myristoyl, propanoyl, or acetyl); R 10 is -C(O)C 1-6 is alkyl (e.g., R 10 is -C(O)C 1-3 Alkyl, for example, -C(O)CH3.

[0054] In some embodiments, the compounds of Formula (I), (II), (Ia) and / or (Ib) have the stereochemistry shown in Formula (III) below: [ka]

[0055] In some embodiments, the epoxide at the 6,7 positions is above the plane of the ring system. In other embodiments, the epoxide at the 6,7 positions is below the plane of the ring system. In some embodiments, the R2 group at the 12 position is S, and in other embodiments, the R2 group at the 12 position is R.

[0056] In some embodiments, the epoxy tigliane compound of formula (III) is a compound of formula (IIIa): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is hydrogen and C 1-6 alkyl (e.g., R is selected from C 1-3 alkyl, e.g., CH3, etc.); R2 is -OC(O)C 1-7 Alkyl, -OC(O)C 2-7 Alkenyl and -OC(O)C 2-7 alkynyl (e.g., R2 is -O-tigloyl, -O-2-methylbutanoyl, -O-hexanoyl, -O-myristoyl, or -O-propanoyl); R3 is -OC(O)C 1-7Alkyl, -OC(O)C 2-7 Alkenyl and -OC(O)C 2-7 alkynyl (e.g., R2 is -O-tigloyl, -O-2-methylbutanoyl, -O-hexanoyl, or -O-acetyl); R4 and R5 are independently hydrogen and C 1-6 alkyl (e.g., R and R are independently selected from C 1-3 alkyl, e.g., CH3, etc.); R6 is hydrogen or -C(O)C 1-6 alkyl (e.g., R6 is -C(O)C 1-3 alkyl, such as -C(O)CH3; R7 is hydroxy; and R8 is hydrogen or C 1-6 alkyl (e.g., R is C 1-3 alkyl, for example, CH3.

[0057] In some embodiments, the epoxy tigliane compound of formula (III) is a compound of formula (IIIb): [ka] or a pharma- ceutically acceptable salt thereof; During the ceremony, R1 is C 1-6 alkyl (e.g., R is selected from C 1-3 alkyl, e.g., CH3, etc.); R2 is -OC(O)C 1-7 Alkyl and -OC(O)C 2-7 alkenyl (e.g., R2 is -O-tigloyl, -O-2-methylbutanoyl, -O-hexanoyl, -O-myristoyl, or -O-propanoyl); R3 is -OC(O)C 1-7 Alkyl and -OC(O)C 2-7 alkenyl (e.g., R2 is -O-tigloyl, -O-2-methylbutanoyl, -O-hexanoyl, or -O-acetyl); R4 and R5 are independently C 1-6 alkyl (e.g., R and R are independently 1-3 alkyl, e.g., CH3, etc.); R6 is hydrogen or -C(O)C 1-6 alkyl (e.g., R6 is hydrogen or -C(O)C 1-3 alkyl, such as -C(O)CH3; R7 is hydroxy; and R8 is C 1-6 alkyl (e.g., R is C 1-3 alkyl, for example, CH3.

[0058] In certain embodiments, the epoxytigliane compound is selected from the following: 12-Tigloyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 1); 12,13-di-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 2); 12-Hexanoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 3); 12,13-Dihexanoyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 4); 12-Myristoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 5); 12-Tigloyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13-pentahydroxy-20-acetyloxy-1-tiglien-3-one (compound 6); 12-Myristoyl-13-acetyloxy-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 7); 12-propanoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 8); 12,13-ditigloyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 9); and 12-(2-Methylbutanoyl)-13-tigloyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one (compound 10).

[0059] In some embodiments, the epoxy tigliane compound is tigilanol tiglate (abbreviated "TT"), which is (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoethyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexyl sahydroxy-1-tigliane-3-one (also known as [(1R,2S,4R,5S,6S,10S,11R,12R,13R,14S,16R)-5,6,11-trihydroxy-4-(hydroxymethyl)-8,12,15,15-tetramethyl-13-[(E)-2-methylbut-2-enoyl]oxy-7-oxo-3-oxapentacyclo[9.5.0.0 2,4 .0 6,10 .0 14,16 ]hexadec-8-en-14-yl](2S)-2-methylbutanoate; the chemical name is calculated by Lexichem TK 2.7.0 (PubChem release 2021.05.07) and has the following structure: [ka]

[0060] Second Therapeutic Agent The second therapeutic agent can be a cancer chemotherapeutic agent or radiation. In some embodiments, the second therapeutic agent is a chemotherapeutic agent. In other embodiments, the second therapeutic agent is radiation.

[0061] Without wishing to be bound by theory, it is hypothesized that the epoxytigliane compounds of formula (I) may be synergistic with cancer therapies that damage DNA or inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion. Thus, the second therapeutic agent is one that damages DNA, causing cell death, or kills tumor-associated host-derived cells, such as myeloid-derived suppressor cells (MDSCs), that support tumor cell proliferation and / or invasion.

[0062] Examples of agents that damage DNA include DNA intercalators (substances that insert into the DNA structure of a cell and bind to DNA, causing DNA damage) and DNA alkylating agents.

[0063] In some embodiments, the second therapeutic agent is a chemotherapeutic agent selected from: i) platinum-based chemotherapy agents, in particular cisplatin and carboplatin, more particularly cisplatin (Dasari and Tchounwou, 2013; Bracci et al., 2014); ii) DNA intercalators or inhibitors of the enzyme topoisomerase II, in particular doxorubicin or mitoxantrone (Cheung-Ong et al., 2013); iii) DNA alkylating agents, such as cyclophosphamide ( Cheung-Ong et al., 2013 ; Hall and Tilby, 1992 ); iv) antimetabolites, such as gemcitabine or fluorouracil, in particular 5-fluorouracil (Cheung-Ong et al., 2013; Albeituni et al., 2013); v) Vinca alkaloids, e.g., vincristine ( Moudi et al., 2013 ; Mohammadgholi et al., 2013 ); vi) compounds that bind to GC-rich DNA, such as bleomycin; and vii) MDSC inhibitors, such as phosphodiesterase-5 inhibitors, e.g., ludenafil; amiloride; or COX-2 and PEG-2 inhibitors such as celecoxib (Albeituni et al., 2013).

[0064] In some embodiments, the second therapeutic agent is or comprises cisplatin or carboplatin, hi some embodiments, the second therapeutic agent is or comprises cisplatin.

[0065] In some embodiments, the second therapeutic agent is or comprises 5-fluorouracil.

[0066] In some embodiments, the second therapeutic agent is or includes doxorubicin, which is believed to damage DNA through intercalation or direct alkylation, potentially resulting in the formation of double-stranded breaks in DNA.

[0067] In some embodiments, the second therapeutic agent is irradiation. Irradiation can occur, for example, by external beam radiation therapy (a high-energy beam is delivered to the site of the cancer from a machine outside the body) or brachytherapy (a beam is delivered from a radiation source placed in an adjacent site or inside the body). At high doses, radiation therapy (or "radiotherapy") kills cancer cells and / or slows their growth by damaging DNA. The radiation source can be any radioactive material commonly used in cancer radiation therapy. In some embodiments, the radiation source is cesium (e.g., cesium-137). In some embodiments, the radiation source is cobalt (e.g., cobalt-60).

[0068] In some embodiments, the treatment method includes an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and cisplatin. This combination can be used, for example, to treat squamous cell carcinoma (SCC), including in situ SCC and invasive SCC (including UV-induced SCC).

[0069] In some embodiments, the treatment method includes an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tigliane-3-one) and 5-fluorouracil. This combination can be used, for example, to treat squamous cell carcinoma (SCC), including in situ SCC and invasive SCC (including UV-induced SCC).

[0070] In some embodiments, the treatment method includes an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and irradiation. This combination can be used, for example, to treat squamous cell carcinoma (SCC), including in situ SCC and invasive SCC (including ultraviolet light-induced SCC), and adenocarcinoma, including colon adenocarcinoma.

[0071] In some embodiments, the treatment method includes an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and doxorubicin. This combination can be used, for example, in the treatment of melanoma and lung cancer.

[0072] In some embodiments, the treatment method includes an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tigliane-3-one) and irradiation.

[0073] The second chemotherapeutic agent is not an immune checkpoint inhibitor.

[0074] The epoxytigliane and the second therapeutic agent may be administered according to any suitable administration regimen, for example, they may be administered simultaneously, sequentially, or on different occasions.

[0075] In some embodiments, the treatment regimen includes only two components, i.e., the administration of an epoxytigliane compound and a second therapeutic agent. For example, it may include the administration of an epoxytigliane compound and a chemotherapeutic agent that only damages DNA. Alternatively, it may include the administration of an epoxytigliane compound and a chemotherapeutic agent that only inhibits tumor-associated host-derived cells that support tumor cell proliferation and / or invasion. As a further alternative, the treatment regimen may include the administration of an epoxytigliane compound and radiation only.

[0076] In some embodiments, the treatment regimen includes administration of only an epoxytigliane compound and one second therapeutic agent, e.g., one chemotherapeutic agent that damages DNA or one chemotherapeutic agent that inhibits tumor-associated host-derived cells that support tumor cell proliferation and / or invasion.

[0077] In some other embodiments, the treatment regimen includes administration of an epoxytigliane compound and two or more second therapeutic agents (e.g., two, three, four, or five second therapeutic agents), which may include, for example, administering two chemotherapeutic agents that damage DNA, or two chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion, or one each of those two types of chemotherapeutic agents.

[0078] In some embodiments, the treatment regimen does not include administration of a therapeutic agent other than an epoxytigliane compound and a second therapeutic agent. In some other embodiments, the treatment regimen may include administration of an epoxytigliane compound, a second therapeutic agent as defined herein, and one or more (e.g., one, two, three, four, or five) additional therapeutic agents.

[0079] composition Although the epoxytigliane compound or a pharma- ceutically acceptable salt thereof and the second chemotherapeutic agent can be administered as is, it may be more convenient to administer the epoxytigliane compound and the second chemotherapeutic agent, each together with a pharma- ceutically acceptable carrier, diluent and / or excipient, in the form of one or more pharmaceutical compositions.

[0080] The pharmaceutical uses and dosage forms and amounts of the compositions can be readily determined by those skilled in the art.

[0081] In certain embodiments, the epoxytigliane compounds are formulated for administration directly onto or into the tumor being treated. In some embodiments, the epoxytigliane compounds are formulated for local administration in the form of a gel, ointment, lotion, cream, or transdermal patch that is applied directly to the tumor being treated. In other embodiments, the epoxytigliane compounds are formulated for injection, particularly intratumoral injection, where the compound is injected into one or more locations within the tumor.

[0082] The second chemotherapeutic agent can be administered by any means that can deliver the molecule systemically or locally. In some embodiments, the second chemotherapeutic agent is conveniently administered by injection, for example, intravenous, intraarticular, intramuscular, intradermal, subcutaneous or intraperitoneal injection. The second chemotherapeutic agent can also be formulated for local administration by injection, for example, intratumoral injection. Pharmaceutically acceptable carriers and acceptable carriers for systemic or local administration can also be incorporated into the composition of the second chemotherapeutic agent.

[0083] In some embodiments, the epoxytigliane compound and the second chemotherapeutic agent are administered separately, simultaneously or sequentially, hi other embodiments, the epoxytigliane compound and the second chemotherapeutic agent are administered in a single composition, e.g., a single composition suitable for intratumoral administration or a single composition formulated for systemic administration.

[0084] In another embodiment, a pharmaceutical composition comprising an epoxytigliane compound or a pharma- ceutically acceptable salt thereof and a second chemotherapeutic agent is provided, optionally with one or more pharma- ceutically acceptable carriers.

[0085] Suitably, the pharmaceutical composition comprises a pharma- ceutically acceptable excipient or acceptable vehicle. By "pharma-ceutically acceptable excipient" is meant a solid or liquid filler, diluent or encapsulating substance that can be safely used. Depending on the particular route of administration, various carriers well known in the art can be used. These carriers or excipients may be selected from the group including sugars, starches, cellulose and its derivatives, cyclodextrins, malt, gelatin or other gelling agents, polymers, talc, calcium sulfate, vegetable oils, synthetic oils, alcohols and / or polyols, alginic acid, phosphate buffers, emulsifiers, isotonic saline, and sterile water.

[0086] Liquid formulations include solutions, suspensions, and emulsions, such as water or water-propylene glycol solutions. For example, liquid formulations for parenteral injection can be formulated as aqueous solutions of 1,2-propanediol, dimethylsulfoxide (DMSO), gamma cyclodextrin or 2-hydroxypropyl-β-cyclodextrin, saline, or polyethylene glycol, with or without buffering agents. The preferred pH range is 3.0-4.5. Suitable buffers buffer the formulation at pH 3.5-4.5 and include, but are not limited to, acetate buffer and citrate buffer.

[0087] The composition of the epoxytigliane compound and / or the second chemotherapeutic agent may be formulated for parenteral administration (e.g., by injection, e.g., bolus injection or continuous infusion), and may be provided in unit dose form in ampoules, prefilled syringes, small volume infusions, or multi-dose containers with added preservatives. The composition may take the form of a suspension, solution, gel, or emulsion in an oily or aqueous vehicle, and may contain formulatory agents, such as suspending, stabilizing, and / or dispersing agents. Alternatively, the active ingredient may be in powder form, obtained by aseptic isolation of a sterile solid or lyophilization from solution, so that it can be mixed with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

[0088] Pharmaceutical compositions of the epoxytigliane compound and / or second chemotherapeutic agent suitable for administration may be provided in discrete units such as syringes, vials, tubes or sachets, each containing a predetermined amount of one or more pharma- ceutically active compounds or extracts of the invention, as a powder or granules, or as a solution or suspension in an aqueous solution, a cyclodextrin solution, a non-aqueous solution, an oil-in-water emulsion or a water-in-oil emulsion, or as a solution or suspension in a cream or gel, or as a suspension of microparticles or nanoparticles including the epoxytigliane compound, including but not limited to silica or polylactic acid microparticles or nanoparticles. Such compositions may be prepared by any method of pharmacy, but all methods include the step of bringing into association one or more pharma-ceutically active compounds of the invention with the carrier which constitutes one or more more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the agent of the invention with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired form.

[0089] For local administration to the epidermis or other organs, the epoxytigliane compound and / or the second chemotherapeutic agent can be formulated as a gel, ointment, emulsion, paste, cream or lotion, or as a transdermal patch. Gels can be prepared using a suitable thickening agent, which is added to the aqueous / alcoholic composition of the compound. Suitable thickening or gelling agents are well known in the art, such as Carbomer 940, a polyvinyl carboxy polymer. Ointments and creams can be formulated, for example, with an aqueous or oily base with the addition of a suitable thickening and / or gelling agent. Lotions can be formulated with an aqueous or oily base, and will generally also contain one or more emulsifiers, stabilizing agents, dispersing agents, suspending agents, thickening agents, or coloring agents.

[0090] Formulations suitable for topical administration also include solutions or suspensions which can be applied topically in the form of a bath or soak, or a spray, or can be absorbed into a dressing.

[0091] When the second chemotherapeutic agent is a small molecule, it can be administered by any suitable means, including oral, topical, rectal, parenteral, sublingual, buccal, intravenous, intraarticular, intramuscular, subcutaneous, inhalation, intraocular, intraperitoneal, intraventricular, transdermal, etc., as is known in the pharmaceutical arts.

[0092] Dosing regimen In some embodiments, the epoxytigliane compound is administered in the same composition as the second chemotherapeutic agent. However, in certain embodiments, the second chemotherapeutic agent is administered in a separate composition from the epoxytigliane compound. If the second therapeutic agent is radiation, it will, of course, be administered separately from the administration of the epoxytigliane compound.

[0093] In some embodiments, the epoxytigliane compound is administered directly to the tumor, for example, by topical administration or intratumoral injection.

[0094] In some embodiments, the epoxy tigliane compound is administered once to the tumor. In other embodiments, the treated tumor is monitored and further administration of the epoxy tigliane compound may be required if the tumor does not respond adequately to treatment. In embodiments in which the tumor is treated locally, the epoxy tigliane compound may be administered multiple times over a period of time, for example, daily for one week or once a week for 4-10 weeks. One skilled in the art of monitoring a treated subject will be able to determine an appropriate administration schedule, which will vary depending on the response to treatment.

[0095] In certain embodiments, the epoxytigliane compound is administered only once by intratumoral injection.

[0096] In some embodiments, the amount of the epoxytigliane compound administered is a sub-therapeutic amount.

[0097] In some embodiments, the second chemotherapeutic agent is administered at least once before, simultaneously, or consecutively with the epoxy tigliane compound. In some embodiments, the second chemotherapeutic agent is administered once before, simultaneously, or after administration of the epoxy tigliane compound. For example, in some embodiments, the second chemotherapeutic agent is administered within 24 hours before, immediately before, simultaneously, immediately after, or within 24 hours after administration of the epoxy tigliane compound, particularly immediately before, simultaneously, or immediately after administration of the epoxy tigliane compound. By immediately before or immediately after, it is meant that administration of the second chemotherapeutic agent occurs less than 2 hours, particularly less than 1 hour, and more particularly less than 30 minutes before or after administration of the epoxy tigliane compound.

[0098] In some embodiments, multiple doses of the second chemotherapeutic agent are administered beginning prior to or simultaneously with administration of the epoxytigliane compound and continuing for a period of time after administration of the epoxytigliane compound. In some embodiments, the second chemotherapeutic agent is administered multiple times periodically before and after administration of the epoxytigliane compound. In certain embodiments, the second chemotherapeutic agent is administered prior to administration of the epoxytigliane compound, sequentially or simultaneously with administration of the epoxytigliane compound, and at least once after administration of the epoxytigliane compound. For example, the second chemotherapeutic agent may be administered 24 hours prior to administration of the epoxytigliane compound, then the second chemotherapeutic agent may be administered sequentially or simultaneously with the epoxytigliane compound, or immediately prior to or immediately after administration of the epoxytigliane compound, and thereafter the second chemotherapeutic agent may be administered one or more times for 1 to 3 months after administration of the epoxytigliane compound, for example, once per week, once every 5 days, once every 4 days, once every 3 days, once every 2 days or daily, particularly once every 1 to 3 days, and more particularly once every 2 days. Subsequent administrations of the second chemotherapeutic agent may be administered 1 to 10 times the second chemotherapeutic agent after administration of the epoxytigliane compound. In particular, the administration may be continued for 1 to 8 doses, 1 to 6 doses, 1 to 4 doses, 1 to 3 doses, or 1 to 2 doses.

[0099] In some embodiments, the epoxytigliane compound is administered in an effective amount. By "effective amount" is meant the amount necessary to at least partially achieve the desired response, e.g., reducing the size of the tumor and completely destroying the tumor. The amount will vary depending on the health and physical condition of the individual being treated, the taxonomic group of the individual being treated, the formulation of the composition, the size of the tumor, the evaluation of the medical condition, the second therapeutic agent being used in combination, and other relevant factors. It is expected that the amount will fall within a relatively broad range that can be determined by routine testing. An effective amount is, for example, an amount that reduces the tumor volume by 1 cm 2 per administration. 3 The dosage may range from about 0.1 μg to 20 mg per cm of tumor volume per administration. 3For example, one dose is administered to treat a tumor volume of 1 cm 3 In one embodiment, when the dose is administered intratumorally, the dose is in the range of 0.5 μg to 5 mg per cm of tumor volume. 3 0.01 to 1 mg per cm of tumor volume, for example 3 0.1-1 mg per tumor volume of 1 cm 3 0.1-0.7 mg per cm2 tumor volume 3 0.1-0.8 mg per cm of tumor volume, for example 3 In another embodiment, the dosage is in the range of 0.001 mg to 20 mg per administration, for example, 0.005 mg to 15 mg per administration, particularly, 0.05 mg to 10 mg per administration, and more particularly, about 0.1 mg to about 5 mg per administration. The dosing regimen may be adjusted to obtain an optimal therapeutic response. For example, in some embodiments, if the administration is intratumoral, the epoxytigliane compound is administered once and the progress of the treatment is monitored. In some embodiments, if the tumor does not completely resolve or if the tumor recurs, a second dose may be administered. In some embodiments, if the administration is local, the topical compound formulation may be administered directly to the tumor site in the form of a gel, cream, ointment, or lotion. The frequency of treatment depends on the tumor, its size, the subject being treated, etc. In some embodiments, the topical formulation may be applied weekly until the tumor has disappeared. In other embodiments, the treatment is a monotherapy and a second treatment is administered only if the tumor does not disappear completely.

[0100] In some embodiments, the epoxytigliane compound is administered in a subtherapeutic amount. A subtherapeutic amount is an amount that is less than would be expected to be effective if the epoxytigliane compound were administered alone. For example, a subtherapeutic amount can be administered in ... or equal to 1 cm of tumor volume. 3 The effective amount may be at the lower end of the above-mentioned effective amounts, 0.1 to 0.2 mg per dose.

[0101] The second chemotherapeutic agent may also be administered in an effective amount. Again, the amount of the second chemotherapeutic agent that is deemed effective will depend on the subject, their health and physical condition, the identity of the second chemotherapeutic agent, the composition formulation, and an evaluation of the medical situation. It is expected that the amount of the second chemotherapeutic agent will fall within a fairly wide range of amounts. An effective amount may be in the range of about 0.1 ng to about 500 mg per kg of body weight, 100 μg to 100 mg per kg of body weight, 1 mg to 50 mg per kg of body weight, or 1 mg to 20 mg per kg of body weight. In another embodiment, the actual dosage may be in the range of 1 μg to 1 g, e.g., 100 μg to 750 mg per administration. An effective amount may be sub-therapeutic when used as a monotherapy, but is effective when used in combination with an epoxytigliane compound.

[0102] When the second therapeutic agent is radiation, e.g., external beam radiation, the radiation can be administered at least once. In some embodiments, the radiation is administered only once, either within 24 hours before, immediately before, immediately after, or within 24 hours after administration of the epoxy tigliane compound, particularly immediately before or immediately after administration of the epoxy tigliane compound. Immediately before or immediately after means that the radiation occurs less than 2 hours before or less than 2 hours after administration of the epoxy tigliane compound, particularly less than 1 hour, and more particularly less than 30 minutes.

[0103] In some embodiments, radiation is administered multiple times, for example, radiation can be administered 24 hours prior to administration of the epoxy tigliane compound, and then administered sequentially or simultaneously with the epoxy tigliane compound, either immediately prior to or immediately following administration of the epoxy tigliane compound, and then one or more doses after administration of the epoxy tigliane compound, for example, once per week, once per 5 days, once per 4 days, once per 3 days, once per 2 days, or every day.

[0104] In some embodiments, the amount of radiation administered is 1-8 Gray (Gy) per infusion. In certain embodiments, the amount of radiation administered is 1-6 Gy, 2-4 Gy, or about 2 Gy per infusion, for example, to administer a total dose of 8 Gy-50 Gy. In some embodiments, radiation is administered once at a suboptimal dose, for example, 2 Gy-8 Gy, particularly 2 Gy-4 Gy, or 1 Gy-2 Gy total dose.

[0105] Subjects that may be treated with the combination therapy are mammals, birds, aquatic animals such as fish, or reptiles. In some embodiments, the subject is a human, an experimental animal, such as a mouse, rat, or rabbit, a companion animal, such as a dog or cat, a working animal, such as a horse, donkey, a livestock animal, such as a cow, a cattle, a pig, a sheep, a goat, a deer, a llama, an alpaca, or a captive wild animal, such as one in a zoo or wildlife park, including a lion, a leopard, a cheetah, an elephant, a zebra, antelope, a giraffe, a koala, a kangaroo, and a reptile, such as a crocodile, a lizard, a snake, a bird, particularly a captive bird, such as a budgerigar or a canary, a parrot, a cockatoo, a parrot, or a fish, particularly a captive fish, such as a tropical fish (zebrafish, guppy, Siamese fighting fish, clownfish, cardinal tetra, etc.), a dolphin, a whale, etc. In certain embodiments, the subject is a human or a companion animal.

[0106] In some embodiments, the subject is immunocompetent. In other embodiments, the subject is immunosuppressed or immunodeficient.

[0107] kit The epoxytigliane compound and the second chemotherapeutic agent compositions may be formulated separately and sold together as a kit or package, each kit containing a dose of each compound to achieve treatment of the tumor.

[0108] In some embodiments, the epoxytigliane composition is formulated for topical administration, such as, for example, a gel, lotion, cream, or ointment, or impregnated into a bandage. In other embodiments, the epoxytigliane compound is formulated for injection, such as, for example, intratumoral injection. In this embodiment, the epoxytigliane formulation may be present in the kit as a liquid that can be taken up into a syringe, as a powder or solid formulation that is solubilized in a carrier prior to injection, or as a pre-filled syringe.

[0109] Each kit may include one or more doses of the epoxytigliane compound. In one embodiment, the kit includes a single dose of the epoxytigliane compound in a formulation suitable for intratumoral injection. In another embodiment, the kit includes a topical formulation of the epoxytigliane compound that includes multiple doses for application to a tumor.

[0110] In some embodiments, the second chemotherapeutic agent is formulated for parenteral administration in a single bolus dose or multiple dose form.For example, the kit may include the second chemotherapeutic agent in a pre-filled syringe, either as a liquid in a vial ready to be taken into a syringe, or as a solid ready to be dissolved before being taken into a syringe.The liquid or solid formulation may be a single dose formulation or a multiple dose formulation.Alternatively, the kit may include multiple doses of the second chemotherapeutic agent, each of which may be separately formulated in a pre-filled syringe, either as a liquid in a vial ready to be taken into a syringe, or as a solid ready to be dissolved and taken into a syringe.

[0111] The kit may optionally further include an insert containing instructions for use of each formulation, including how to prepare each dose, how to administer each dose, and when to administer each dose.

[0112] In some embodiments, the kits include an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and can be used for irradiation to treat tumors, such as squamous cell carcinoma (SCC), including in situ SCC and invasive SCC, and adenocarcinoma, including colon adenocarcinoma.

[0113] In some embodiments, the kit comprises an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and cisplatin. The kit can be used, for example, to treat squamous cell carcinoma (SCC), including in situ SCC and invasive SCC.

[0114] In some embodiments, the kit comprises an epoxytigliane compound (e.g., (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and 5-fluorouracil. The kit can be used, for example, to treat squamous cell carcinoma (SCC), including in situ SCC and invasive SCC.

[0115] In some embodiments, the kit includes an epoxytigliane compound (e.g., (,4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoatyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one) and doxorubicin. This kit can be used, for example, to treat melanoma and lung cancer.

[0116] The present disclosure is further illustrated by the following non-limiting examples. EXAMPLES

[0117] Example 1: Combination of Compound 1 and Cisplatin in vivo Ethical approval was obtained from the Queensland Institute of Medical Research (QIMR) Animal Ethics Committee prior to commencing in vivo experiments. All mice were kept under pathogen-free conditions in the QIMR animal care facility, housed in cages with filtered air available, and food and water available ad libitum. Mice were handled and procedures performed using aseptic technique in a laminar flow hood.

[0118] Cancer cells were seeded in T175 tissue culture flasks (Thermo Scientific) to expand cell numbers prior to inoculation into subject mice. Upon reaching 70-80% confluence, cells were trypsinized and resuspended in RPMI-1640 medium for collection, and cells were counted using a hemocytometer. Cell volumes were centrifuged and pellets were reconstituted with RPMI-1640 medium to a volume sufficient for the number of inoculations injected at two sites per mouse subject, requiring 100 μL per injection. Cell suspensions were kept on ice and transported to the animal facility for injection into subjects.

[0119] Details of the mice used and the cell lines used in the combination experiments are provided in Table 1. BALB / c Foxn1 nuMice were obtained from the Western Australian Animal Resource Centre. C3H / HeNCr mice were bred in the QIMR animal facility. [Table 1]

[0120] C3H / HeNCr mice were shaved the day before inoculation (Wahl, Illinois, USA). Restrained subjects were inoculated on both the left and right hindquarters by subcutaneous injection of 100 μL of the appropriate cell suspension using a 27G×1 / 2 "needle" from Terumo® (Tokyo, Japan). Subjects were ear-tagged for identification. After inoculation, mice were monitored twice weekly for tumor development and health, and subjects were monitored daily once tumors developed.

[0121] 30 BALB / c Foxn1 nu Groups of 21 C3H / HeNCr mice and 21 C3H / HeNCr mice, each with 2 tumors per mouse, were divided into 6 experimental groups as shown in Table 2. 30 μg of Compound 1 was administered to 100 mm 3 The doses of 5 and 10 μg were selected as suboptimal doses because they are considered to be therapeutic doses for certain tumors. [Table 2]

[0122] The body weight of each subject was recorded before and after treatment, and at cull. The width and length of each tumor was measured using 150 mm digital Vernier callipers (Kinchrome, Qld, Australia), and individual tumor volumes (mm 3 ) was calculated using the following formula: (Length x (Width 2 ))÷2

[0123] Tumor volume is 70-100mm 324 hours before the expected onset of pulmonary hypertension, test subjects were administered either 2.5 mg / kg cisplatin (0.5 mg / mL in 0.9% saline) or an equivalent volume of 0.9% saline intraperitoneally using a Terumo® 0.5 mL U-100 insulin 29G×1 / 2 “needle”. IP injections were performed by first swabbing the subject’s anterior abdominal wall with an alcohol swab before scruffing the symmetrical dorsal skin and inverting the subject with head down. The 29G needle was used with the bevel upwards to inject into the right lower abdominal quadrant.

[0124] 70~100mm 3 Once the tumors reached a volume of 10 μg / mL, test tumors were injected IT with 50 μL of either 5 μg or 10 μg of Compound 1, or 50 μL of 40% propylene glycol (PG) for the control group.

[0125] Test subjects also received IP injections of cisplatin and 0.9% saline 7 and 14 days after the first injection. Compound 1 concentrations were confirmed by high performance liquid chromatography (HPLC) prior to use.

[0126] Study subjects were observed daily and monitored for up to three months after treatment. Tumor volumes were recorded as described above.

[0127] Total tumor volume was 1000 mm per mouse. 3 Subjects were euthanized by CO2 inhalation when they reached 0.01 mg / kg, if ill, and at the end of the study.

[0128] Data were analyzed with Microsoft Excel and Prism 7, and mean tumor volumes and standard deviations (SD) were calculated. Individual and mean tumor volumes and Kaplan-Meier survival curves were graphed against time of treatment.

[0129] Heterogeneous Models BALB / c Foxn1 nu For the combination of Compound 1 and cisplatin in the mouse model, the individual and mean SCC-15 tumor volumes are shown in Figures 1a and 1b, respectively. Kaplan-Meier survival curves for the xenogeneic model are shown in Figure 2.

[0130] BALB / c Foxn1 nu The mouse model is an immunocompromised host that lacks an intact adaptive immune system. This model allows for the evaluation of Compound 1 / cisplatin combination therapy, in which the host immune system is unlikely to contribute to any therapeutic effect.

[0131] From Figures 1a / b and 2, there is a clear trend in the individual and mean tumor volumes and Kaplan-Meier survival curves that the combination therapy has a superior effect, especially at the higher concentrations of compound 1, i.e., cisplatin IP 2.5mg / kg and compound 1 IT 10μg. This combination resulted in a long-lasting response, with one tumor remaining cured after 60 days. The lower dose of compound 1 (5μg) was not as effective as the higher dose, but still showed a superior effect to monotherapy or no treatment.

[0132] Subject body weight (g) served as a marker for the systemic toxicity of IP cisplatin, allowing us to evaluate whether combination therapy with subtherapeutic concentrations of IP cisplatin would be effective while avoiding the common side effects associated with its administration. The results are shown in Table 3. [Table 3]

[0133] The results show that there is little difference in body weight before and after treatment, but there is a clear increase in body weight for both the control and cisplatin-treated groups by the end of the study. A slight weight loss was observed after treatment with cisplatin, but the weight was restored by the end of the study. For both saline and cisplatin treatments, the weight gain from pre-treatment to the end of the study was statistically significant, but the weight loss from pre-treatment to post-treatment with cisplatin was not statistically significant.

[0134] Similar Models For the combination of Compound 1 and cisplatin in the C3H / HNCr syngeneic mouse model, the individual and mean UV-13-1 tumor volumes are shown in Figures 3a and 3b, respectively. Kaplan-Meier survival curves for the syngeneic mouse model are shown in Figure 4.

[0135] The C3H / HeNCr mouse model inoculated with UV-13-1 tumors is a syngeneic model in which the mice are immunocompetent and the impact of the immune system on treatment can be evaluated. These mice were not culled at day 60 but were monitored regularly (weekly) for recurrence up to day 200.

[0136] As in the xenogeneic model, it is clear that the combination of compound 1 with IT and IP cisplatin is the most effective treatment, showing significant improvements in individual and mean tumor size, as well as improved long-term survival. Figure 4 shows that the combination treatment results in the best duration of response.

[0137] The initial response to treatment with 10 μg of compound 1 and saline is comparable to the combination of compound 1 and cisplatin. However, by day 18, monotherapy with compound 1 is significantly less effective. The average tumor size also reflects this trend. Only the combination of compound 1 and cisplatin produced a sustained tumor response, with some subjects showing no signs of recurrence at the end of the study. Both 5 μg and 10 μg of compound 1 were more effective than IP cisplatin alone.

[0138] The results of the combination in the syngeneic model were more pronounced and more durable than those in the xenogeneic model: in the xenogeneic model, only one tumor remained cured at day 60 with the combination therapy, whereas in the syngeneic model, nine tumors remained cured at day 200 after treatment.

[0139] In the syngeneic model, weight gain was also used as an indicator of cisplatin toxicity. The results are shown in Table 4. [Table 4]

[0140] The results show that in the saline controls, the change in body weight during the experiment was not statistically significant. The pre- and post-treatment body weight change with the IP cisplatin group was statistically significant (p=<0.0001), but the weight gain from pre-treatment to death in this group was not significant. The statistically significant decrease in body weight from pre-treatment to post-treatment indicates that even with sub-therapeutic concentrations of cisplatin, there is some toxicity.

[0141] summary A summary of survival with each treatment is shown in Table 5. [Table 5]

[0142] Example 2: Combination of Compound 1 and Irradiation In Vivo Ethical approval was obtained from the Queensland Institute of Medical Research (QIMR) Animal Ethics Committee prior to commencing in vivo experiments. All mice were kept under pathogen-free conditions in the QIMR animal care facility, housed in cages with filtered air available, and food and water available ad libitum. Mice were handled and procedures performed using aseptic technique in a laminar flow hood.

[0143] Cancer cells were seeded in T175 tissue culture flasks (Thermo Scientific) to expand cell numbers prior to inoculation into subject mice. Upon reaching 70-80% confluence, cells were trypsinized and resuspended in RPMI-1640 medium for collection, and cells were counted using a hemocytometer. Cell volumes were centrifuged and pellets were reconstituted with RPMI-1640 medium to a volume sufficient for the number of inoculations injected at two sites per mouse subject, requiring 100 μL per injection. Cell suspensions were kept on ice and transported to the animal facility for injection into subjects.

[0144] Details of the mice used and the cell lines used in the combination experiments are provided in Table 6. BALB / c Foxn1 nu and C57BL / 6 mice were obtained from the Western Australian Animal Resource Centre. [Table 6]

[0145] The MC38 cell line is a murine colon adenocarcinoma cell line derived from C57BL / 6 mice (Kerafast, ENH204).

[0146] C57BL / 6 mice were shaved the day before inoculation (Wahl, IL, USA). Restrained subjects were inoculated in both the left and right hindquarters by subcutaneous injection of 100 μL of the appropriate cell suspension using a Terumo® (Tokyo, Japan) 27G×1 / 2 "needle". Subjects were ear tagged for identification. After inoculation, mice were monitored twice weekly for tumor development and health, and subjects were monitored daily once tumors developed.

[0147] 30 BALB / c Foxn1 nu Mice and groups of 30 C56BL / 6 mice, each with 2 tumors per mouse, were divided into 6 experimental groups as shown in Table 7. Since 30 μg of Compound 1 was considered a therapeutic dose, 7.5 μg was selected as the suboptimal dose. [Table 7]

[0148] The width and length of each tumor were measured using a 150 mm digital caliper (Kinchrome, Queensland, Australia) and individual tumor volumes (mm 3 ) was calculated using the following formula: (Length x (Width 2 ))÷2

[0149] 70~100mm 3 Upon reaching a volume of 1000 mg / kg, the tumors under study were irradiated with 0, 5, or 10 Gy. Each mouse was sedated via intraperitoneal injection with a mixture of ketamine (80 mg / kg in an 8 mg / mL solution, Ceva, NSW, Australia) and midazolam (4 mg / kg in a 0.4 mg / mL solution, Sandoz, Hozkirchen, Germany), and proper positioning was ensured in the irradiator (Gammacel 40 Extractor). Mice were double-chambered during transfers to and from the irradiator. During irradiation, mice were restrained with lead shielding to prevent radiation exposure to sites other than the flank, where the tumor was located. Upon return to the animal facility, mice were injected intratumorally (IT) with 50 μL of 7.5 μg of compound 1 or, for the control group, 50 μL of 40% propylene glycol (PG) using a Terumo 0.5 mL U-100 insulin 29G×1 / 2 "needle". At the end of the treatment, the mice were warmed with an electric blanket, placed on their side, and observed until they became mobile.

[0150] Study subjects were observed daily and monitored for up to three months after treatment. Tumor volumes were recorded as described above.

[0151] Total tumor volume was 1000 mm per mouse. 3 Subjects were administered CO 2 The animal was euthanized by inhalation.

[0152] Data were analyzed in Microsoft Excel and Prism 7, and mean tumor volumes and standard deviations (SD) were calculated. Individual and mean tumor volumes and Kaplan-Meier survival curves were graphed against time of treatment.

[0153] Heterogeneous Models BALB / c Foxn1 nu For the combination of Compound 1 and radiation in the mouse model, the individual and mean SCC-15 tumor volumes are shown in Figures 5a and 5b, respectively. Kaplan-Meier survival curves for the xenogeneic model are shown in Figure 6.

[0154] BALB / c Foxn1 nu The mouse model is an immunocompromised host that lacks an intact adaptive immune system. This model allows for the evaluation of compound 1 / irradiation combination therapies, in which the host immune system is unlikely to contribute to any therapeutic effect.

[0155] Individual and mean tumor volumes in Figures 5a and 5b show that the combination of compound 1 with radiation resulted in a significant and sustained reduction in tumor size compared to the other combinations. The combination of 10 Gy and 7.5 μg IT compound 1 provided the best results, with 5 Gy radiation and 7.5 μg IT compound 1 slowing tumor growth. The next best results were seen with either 7.5 μg IT compound 1 or 10 Gy radiation, both of which induced a delayed tumor response. Irradiation with 10 Gy induced a delayed tumor response, showing an initial steady decrease in mean tumor volume before plateauing and regressing slightly between days 20 and 25. Tumors began to recur after day 25, and mean tumor volume began to increase. There were few obvious differences between treatment with 5 Gy radiation alone and untreated controls.

[0156] The findings in Figures 5a and 5b are replicated in the Kaplan-Meier survival curves in Figure 6. Figure 6 shows that the combination of Compound 1 and radiation provides a significant survival benefit compared to monotherapy. In this xenogeneic model, the combination of higher doses of Compound 1 and radiation has a 50% survival rate at day 200, while the combination of Compound 1 and 5 Gy of radiation shows a 20% survival rate. The other treatments show a progressive and steady decline over the first 30 days after treatment.

[0157] Similar Models Individual and mean MC38 tumor volumes for Compound 1 and radiation in the C57BL / 6 syngeneic mouse model are shown in Figures 7a and 7b, respectively. Kaplan-Meier survival curves for the syngeneic mouse model are shown in Figure 8.

[0158] The MC38 tumor-inoculated C57BL / 6 mouse model is a syngeneic mouse model in which the mice are immunocompetent and the impact of the immune system on treatment can be assessed.

[0159] In the syngeneic model, the combination of compound 1 and radiation further results in a significantly improved tumor response, as evidenced by the individual and mean tumor volumes in Figures 7a and 7b. For each compound 1 and radiation combination, there were two tumors that remained cured even 200 days after treatment. All remaining tumors recurred and showed rapid growth 20-50 days after treatment. Despite early recurrences, the outcome of the combination therapy remained much better than the other monotherapy regimens.

[0160] Kaplan-Meier survival analysis in Figure 8 shows that Compound 1 and radiation treatment is associated with better survival than other combinations of treatments. The two combinations of Compound 1 and radiation (5 Gy and 10 Gy) have similar long-term survival, with 20% survival at the end of the experiment, respectively. Each agent used alone appears to have a similar survival rate.

[0161] Irradiation alone, either 5 Gy or 10 Gy, had a 0% cure rate. However, the addition of compound 1 to irradiation resulted in a significant improvement in cure rates. The percentage of tumors that fully responded to compound 1 and irradiation with 5 Gy or 10 Gy was 80% and 70%, respectively, in the xenogeneic model and 100% in the syngeneic model. IT compound 1 at 7.5 μg alone resulted in a 70% cure rate in both mouse models. The cure rate with compound 1 alone was quite high and close to that of the combination therapy, but the main difference between monotherapy and combination therapy is the duration of response. Compound 1 with either 5 Gy or 10 Gy irradiation improved the duration of the therapeutic response by at least 4.6-fold in the xenogeneic model (11 days vs. 51.5 days and 111 days) and 8.7-fold in the syngeneic model (6.2 days vs. 64.6 days and 65.9 days).

[0162] Median survival times with each treatment are shown in Table 8. [Table 8]

[0163] Example 3: In vitro experiments with compound 1 and chemoirradiation Cell lines were investigated in in vitro experiments to evaluate the combination of Compound 1 with cisplatin, 5-fluorouracil, and radiation. The cell lines are shown in Table 9 and include one skin cell line and one oropharyngeal mucosa cell line. [Table 9]

[0164] cell culture Cells were cultured in Roswell Park Memorial Institute (RPMI-1640; CSL Biosciences, Parkville, Victoria, Australia) medium containing 10% (v / v) fetal calf serum (FCS, CSL Biosciences), 60 μg / mL penicillin (CSL Biosciences), 100 μg / mL streptomycin, 1 mM pyruvate, 0.2 mM nicotinamide, and 3 mM 4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), and were incubated at 37°C with 5% carbon dioxide and 95% humidity (Heracell Vios 160i CO2 incubator, Thermo Scientific, MA, USA). Cell lines were assessed for confluence twice weekly and, when approaching 90%, were passaged by removing the medium before washing with phosphate-buffered saline (PBS, 137 mM NaCl, 6.5 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.4, QIMR Media Services) and treating with 0.25% (v / v) trypsin (Invitrogen, CA, USA) and 3.8% (v / v) versine in Dulbecco's salt solution (QIMR Media Services). An appropriate volume of cells, ranging from 1:5 to 1:10, was harvested and the remainder was recultured in fresh RPMI-1640 / FCS.

[0165] Cell lines were authenticated by short tandem repeat (STR) profiling and mycoplasma testing performed at the QIMR analytical facility.

[0166] The number of cells removed during passaging was measured using a hemocytometer (Fuchs Rosenthal Dark Lines; Hirschmann Laboratories, Eberstadt, Germany). Cells were seeded at 3000 cells / well in 100 μL of medium in flat-bottom 96-well cell culture plates (Corning Incorporated, New York, USA). Cells were cultured for 24 h to ensure attachment to the plates before treatment.

[0167] Single-drug exposure Each cell line was exposed to a range of doses of Compound 1 (20 μg / mL in 100% ethanol), cisplatin (50 μM stock in 0.9% saline, Sigma Aldrich, MO, USA), 5-fluorouracil (5-FU, 50 μM stock in dimethylsulfoxide (DMSO, Sigma Aldrich)), and irradiation. Compound 1 concentrations were confirmed by HPLC prior to use. Dose ranges are shown in Table 10. [Table 10]

[0168] For each of compound 1, cisplatin 5, and -FU, medium was aspirated from the superior most row of the 96-well plate before dispensing 200 μL of each drug stock in RPMI-1640 medium formulated to the maximum desired concentration. Half-dilutions were then performed down the plate, leaving the last row as an untreated control. Each cell line also had an ethanol control that was dispensed in a similar manner as the various drug compounds. After 4, 6, or 24 hours, the drugs were washed out with PBS and 100 μL of fresh RPMI-1640 medium was dispensed.

[0169] Each cell line was irradiated with cesium-137 (Gammacell 40 Exactor, Theratronics, Ontario, Canada) at a dose of 0–8 Gy at a rate of 1 Gy / min with 2 Gy intervals. After treatment, the medium was immediately replaced.

[0170] Each cell line was treated with each drug in triplicate and data across the triplicates were averaged.

[0171] Concomitant exposure Cell lines were exposed to combinations of Compound 1 and other drugs across a wide range of doses, as shown in Table 11. Doses were adjusted to induce suboptimal cell death ( <IC 50) which brings about [Table 11]

[0172] Combination experiments of Compound 1 and cisplatin or 5-FU were performed using various dose time courses. To evaluate the effect of dose time courses on therapeutic efficacy, each cell line was exposed to Compound 1 24 hours before, simultaneously with, and 24 hours after treatment with either cisplatin or 5-FU.

[0173] Cell lines treated with a combination of Compound 1 and radiation were first treated with the assigned duration of radiation immediately prior to treatment with Compound 1. In single agent experiments, wells included no control and ethanol-treated controls. After treatment, cells were incubated for 4 hours before washing with PBS and changing the medium.

[0174] Sulforhodamine B proliferation assay After culturing treated cells for 5–7 days, cell viability was measured using a sulforhodamine B (SRB) proliferation assay. This assay quantitatively assesses the degree of protein staining as a surrogate marker of cell proliferation. Cells were washed with PBS and subsequently fixed with methanol-denatured alcohol (Chem Supply, South Australia) for at least 5 min. After fixation, the methanol-denatured alcohol was removed and cells were again washed briefly with water before staining with 50 μL / well of 0.4% SRB solution in 1% acetic acid solution (Sigma Aldrich) for 60 min. This solution was tipped off and cells were quickly washed twice with 1% acetic acid. The assay was completed by administering 100 μL / well of 10 mM Tris base (unbuffered, pH>9, Invitrogen). Absorbance was measured in an enzyme-linked immunosorbent assay (ELISA) reader (VERSA max microplate reader, Molecular Devices, CA, USA) at 564 nm and 5 s of agitation. Cell survival was graphed as a percentage of absorbance in control wells versus dose.

[0175] statistical analysis Data were analyzed using Microsoft Excel (2016, Prism (Version 7.0, Graphpad software 2016), and Combusyn (Version 1.0, Chou & Martin 2005). Means, standard deviations, and Chou-Talalay combination indices (CI) (Chou (2010)) were calculated and used to detect any significant differences between control and test combinations and to evaluate the relationship between Compound 1 and chemoradiotherapy.

[0176] The Chou-Talalay combination theorem is a statistical method to quantify the effect of combining two or more drugs by calculating a number related to 1. The combination effect (CI-Fa) plot graphically depicts the combination index for a range of drug concentrations. Data points below the line (which represents a combination index of 1) suggest a synergistic relationship. The advantage of the CI-Fa plot is that it shows trends across a wide range of effect levels. In this example setup, the x-axis or effect (Fa) directly correlates with cell survival. For example, an effect (Fa) of 0.1 corresponds to 10% cell survival. Analysis was performed using the software program Combusyn.

[0177] result Single agent Using a 4-h exposure time, cisplatin alone had IC values ​​of 3.125–6.25 μM in both cell lines. 50 SCC-15 was close to the lower limit, whereas A431 was somewhat more resistant and had an IC close to the upper limit. 50 had.

[0178] Using a 4-h exposure time, compound 1 alone had an IC of 62.5–200 µM. 50 SCC-15 was close to the lower limit, while A431 was somewhat more resistant and close to the upper limit.

[0179] Using a 4 h exposure time, 5-FU alone had an IC of 12.5–50 µM in both cell lines. 50 A431 was slightly more sensitive and closer to the lower limit, whereas SCC-15 was slightly more resistant and closer to the upper limit. 50 had.

[0180] Exposure to increasing concentrations of irradiation resulted in a linear decrease in cell survival in both cell lines, with IC 50 Both were approximately 4 Gy.

[0181] For single agent experiments, cell lines were maintained in culture before final transfer to 96-well plates. Treatment for combination experiments involved first exposing cells to either cisplatin, 5-FU, or radiation immediately prior to treatment with five different concentrations of compound 1 for 4 hours. Cell survival was determined using the SRB assay after 7 days of culture compared to untreated controls.

[0182] Use in combination The cell survival of the combination of Compound 1 and cisplatin in A431 cells is shown in Figure 9a. It is clear that the addition of Compound 1 to any concentration of cisplatin results in a significant increase in cell death compared to cisplatin alone. The estimated IC of the combination therapy 50 was equal to or less than 1.75 μM. Most concentrations of Compound 1 had little difference in effect, with the exception of 200 μM.

[0183] The cell survival of the combination of Compound 1 and cisplatin in SCC-15 cells is shown in Figure 9b. It is clear that the addition of Compound 1 to any concentration of cisplatin results in a significant increase in cell death compared to cisplatin alone. The estimated IC 50 was equal to or less than 1.75 μM.

[0184] In summary, combined treatment with cisplatin and compound 1 showed a trend towards decreased cell viability in both cell lines with increasing concentrations of cisplatin, regardless of the concentration of compound 1. The addition of compound 1, even at very low concentrations, led to decreased cell viability compared to cells exposed to cisplatin alone. For both cell lines, the IC 50 is the IC of a single drug alone 50 was decreased compared to

[0185] The cell survival of the combination of Compound 1 and 5-FU in A431 cells is shown in Figure 10a. It is clear that the addition of Compound 1 to any concentration of 5-FU results in a significant increase in cells compared to 5-FU alone. The estimated IC 50 was equal to or less than 1.56 μM.

[0186] The cell survival of the combination of compound 1 and 5-FU in SCC-15 cells is shown in Figure 10b. It is clear that the addition of compound 1 to any concentration of 5-FU results in a significant increase in cell death compared to cisplatin alone. The estimated IC of the combination therapy 50 was equal to or less than 12.5 μM.

[0187] In summary, A431 and SCC-15 cells responded similarly with increases in both 5-FU and Compound 1, resulting in decreased cell survival.

[0188] The cell survival of the combination of Compound 1 and irradiation in A431 cells is shown in Figure 11a. It is clear that the addition of Compound 1 to any concentration of irradiation results in a significant increase in cell death compared to Compound 1 alone.

[0189] The cell survival of the combination of compound 1 and radiation in SCC-15 cells is shown in Figure 11b. Exposure to compound 1 and radiation results in a significant decrease in cell survival. The combination is superior to that of compound 1 alone. Increasing exposure to radiation does not significantly improve the effect.

[0190] IC of the combination of compound 1 and irradiation 50 ranged from 0 to 6.26 μM in A431 cells and from 0 to 1.56 μM in SCC-15 cells.

[0191] Order of Administration In combination therapy with Compound 1 and cisplatin, there was little difference between administering Compound 1 24 hours before or after cisplatin in A431 cells, however, co-treatment significantly improved efficacy across all concentrations of Compound 1.

[0192] In combination therapy of Compound 1 and cisplatin, there was little difference in SCC-15 cells between administering Compound 1 24 hours before or after cisplatin, or administering it simultaneously.

[0193] Combination treatment of compound 1 and 5-FU was somewhat variable in efficacy in A431 cells. Co-treatment produced the greatest cell death across the range of compound 1 concentrations. Pre-treatment with compound 1 was the next most effective, with cell survival approaching that of co-treatment at higher concentrations of 5-FU.

[0194] In the combination therapy of Compound 1 and 5-FU, there was little difference in efficacy between administering Compound 1 24 hours before or after 5-FU or administering it simultaneously in SCC-15 cells.

[0195] Combined Effects The combination effect plot for A431 cells is shown in Figure 12. For each concentration of Compound 1, the IC 50 The combination index is shown in Table 12. [Table 12]

[0196] Using the Chou-Talalay combination index method, it was suggested that Compound 1 has a synergistic relationship with cisplatin, 5-FU and irradiation in A431 cells. All data points for the combination have an index <1.

[0197] The combination effect plot for SCC-15 cells is shown in Figure 13, and the IC 50 The combination index is shown in Table 13. [Table 13]

[0198] Using the Chou-Talalay combination index method, results with SCC-15 cells indicate that compound 1 is synergistic with cisplatin, 5-FU and radiation at low concentrations, and at all concentrations when combined with 5-FU, however, some of the combination indices for radiation and compound 1 were consistent with antagonism (CI>1).

[0199] Example 4: Evaluation of the effect of combination chemotherapy with Compound 1 in a mouse cancer model In this example, the chemotherapy drug doxorubicin was combined with compound 1 in a treatment to evaluate its effect on mouse models of cancer, specifically melanoma and lung cancer.

[0200] Show me how cell line B16-F10-OVA (B16-F10 mouse melanoma cell line overexpressing chicken ovalbumin) were cultured in RPMI-1640 supplemented with 10% FCS (complete medium) and 3 mM HEPES. LLC (Lewis Lung Carcinoma) cells were cultured in DMEM (Gibco) supplemented with 10% FCS. Both cell lines were cultured at 37°C and 5% CO2 in a humidified incubator. Routine testing for mycoplasma was assessed using Myco-Alert (Lonza) and was always negative.

[0201] Animal testing C57BL / 6 mice were purchased from the Animal Resource Centre (ARC) in Murdoch, Western Australia. All animal procedures were carried out in accordance with the NHMRC guidelines (Australian Code for the Care and Use of Animals for Scientific Purposes 8). th The study was approved by the QIMR Berghofer Animal Ethics Committee under approval number A2109-621 in accordance with the National Health and Medical Research Council of Australia (Eds. 2013 Edition; National Health and Medical Research Council of Australia). All mice were housed in specific pathogen-free (SPF) facilities with a 12-h light / dark cycle and had access to food and water at all times.

[0202] Doxorubicin combination therapy C57BL / 6 mice (6–7 weeks old) were inoculated with B16-F10-OVA mouse melanoma cells (2 × 10 per site in 50 μl). 5 cells) or LLC mouse lung cancer cells (5 × 10 per site in 50 μl 5 The tumors were grown to approximately 75–100 mm 3 The tumors were allowed to grow until 10 days after tumor growth. Mice were then dosed intraperitoneally with doxorubicin in saline (3 mg / kg; 60 μg / dose, saline for vehicle). The next day, tumors were injected intratumorally with either vehicle (40% PG / 30 mM acetate) or 5 or 10 μg of compound 1 (50 μl [5 μg] of 100 μg / ml or 50 μl [10 μg] of 200 μg / ml) in 40% PG / 30 mM acetate, pH 4.30, as previously published (Cullen et al., 2021). Treatment with chemotherapy agents was repeated 7 and 14 days later. For the B16-F10-OVA model, 5 mice per group (10 tumors) were used for two experiments (total of 10 mice per group, total of 20 tumors in both experiments). In the LLC tumor model, five mice per group were used per experiment (10 tumors).

[0203] Volume was recorded using digital calipers and measured in mm according to the formula A × b × b × 0.5. 3 The mean tumor volume was expressed as A = tumor length, b = tumor width. Mice were also evaluated for clinical signs of distress during the experiment according to an approved clinical score sheet. Mice were discontinued if they reached an unacceptable clinical score (>6) or if their cumulative tumor burden exceeded 1,000 mm 3 If the tumor growth rate exceeded 100%, the experiment was terminated. All mice that were tumor-free with the above experimental regimen were monitored for an additional 2 months to confirm cure or tumor recurrence / metastasis. At the end of the experiment, the mice were humanely euthanized by CO2 asphyxiation.

[0204] result Treatment of B16-F10-OVA (FIGS. 14a and 14b) tumors in C57BL / 6 mice with either intraperitoneal doxorubicin or saline in combination with intralesional injection of 40% PG / 30 mM acetate had no effect on tumor growth. As expected, treatment with either 5 μg or 10 μg of Compound 1 delayed tumor growth and extended overall survival compared to vehicle alone (p<0.0001, not shown in FIG. 14b). Furthermore, in the B16-F10-OVA model, treatment with doxorubicin in combination with 10 μg of Compound 1 affected tumor growth and extended the time to experimental endpoints when compared to doxorubicin alone and, to a lesser extent, 10 μg of Compound 1 in combination with saline vehicle. In the doxorubicin / 10 μg compound 1 group, 7 of 20 tumors were completely eliminated, whereas in the saline / 10 μg compound 1 group, 4 of 20 tumors were completely eliminated.

[0205] Treatment of LLC (Figures 15a and 15b) tumors in C57BL / 6 mice with either intraperitoneal doxorubicin or saline in combination with intralesional injection of 40% PG / 30 mM acetate had a modest effect on tumor growth delay and survival extension. Furthermore, treatment with either 5 μg or 10 μg of Compound 1 delayed tumor growth and extended overall survival compared to vehicle alone (p<0.0001, not shown in Figure 15b). Furthermore, in the LLC model, treatment with doxorubicin in combination with 10 μg of Compound 1 did not affect tumor growth, resulting in no tumor growth in 3 out of 10 cases. This led to an extension of the time to experimental endpoints when compared to doxorubicin alone and, to a lesser extent, 10 μg of Compound 1 in combination with saline vehicle.

[0206] Consideration Combination treatment with doxorubicin in combination with 10 μg of Compound 1 affected tumor growth delay and overall survival in C57BL / 6 mice bearing B16-F10-OVA or LLC tumors.

[0207] It will be appreciated by those skilled in the art that numerous variations and / or modifications can be made to the above-described embodiments without departing from the broad general scope of the present disclosure, and the present embodiments are therefore to be considered in all respects as illustrative and not restrictive.

[0208] reference Sabrin Husein Albeituni, Chuanlin Ding and Jun Yan.Hampering the Immune Suppressors: Therapeutic Targeting of Myeloid-Derived Suppressor Cells(MDSC) in Cancer.Cancer J.2013;19(6):490-501. Boyle GM,D’Souza MM,Pierce CJ,Adams RA,Cantor AS,Johns JP,Maslovskaya L,Gordon VA,Reddell PW,Parsons PG.Intra-Lesional Injection of the Novel PKC Activator EBC-46 Rapidly Ablates Tumors in Mouse Models.PLoS One 9(10):e108887.https: / / doi.org / 10.1371 / journal.pone.0108887. L Bracci,G Schiavoni,A Sistigu and F Belardelli.Immune-based mechanisms of cytotoxic chemotherapy:implications for the design of novel and rationale-based combined treatments against cancer.Cell Death and Differentiation(2014)21,15-25. Chou TC,Drug combination studies and their synergy quantification using the Chou-Talalay method.2010,Cancer Research,70:440-446. Kahlin Cheung-Ong,Guri Giaever,Corey Nislow DNA-Damaging Agents in Cancer Chemotherapy:Serendipity and Chemical Biology.Chemistry and Biology20(5),648-659,2013. Shaloam Dasari and Paul Bernard Tchounwou.Cisplatin in cancer therapy:molecular mechanisms of action.Eur J Pharmacol.2014 Oct 5;740:364-378. Hall AG,Tilby MJ.Mechanisms of action of,and modes of resistance to,alkylating agents used in the treatment of haematological malignancies.Blood Reviews.(1992)6(3):163-73. Azadeh Mohammadgholi,Azra Rabbani-Chadegani,and Sodabeh Fallah Mechanism of the Interaction of Plant Alkaloid Vincristine with DNA and Chromatin:Spectroscopic Study.DNA Cell Biol.2013 May;32(5):228-235. Maryam Moudi,Rusea Go,Christina Yong Seok Yien,and Mohd.Nazre. Vinca Alkaloids.Int J Prev Med.2013 Nov;4(11):1231-1235. Cullen JK,Boyle GM,Yap PY,Elmlinger S,Simmons JL,BroitN, Johns J,Ferguson B,Maslovskaya LA,Savchenko AI,Mirzayans PM,Porzelle A,Bernhardt PV,Gordon VA,Reddell PW,Pagani A,Appendino G,Parsons PG,Williams CM.Activation of PKC supports the anticancer activity of tigilanol tiglate and related epoxytiglianes.Sci Rep.2021 Jan 8;11(1):207.doi:10.1038 / s41598-020-80397-9.PMID:33420238.

Claims

1. 1. A method of treating a tumor comprising administering a combination of an epoxytigliane compound and a second therapeutic agent, the epoxytigliane compound being a compound of formula (I): 【Chemistry 1】 or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof, During the ceremony, R １ is hydrogen or C １－６ is alkyl; R ２ is -OH or -OR ９ and R ３ is -OH or -OR ９ and R ４ and R ５ are independently hydrogen and C １－６ alkyl; R ６ is hydrogen or -R １０ and R ７ is hydroxy or OR １０ and R ８ is hydrogen or C １－６ is alkyl; R ９ is -C １－２０ Alkyl, -C ２－２０ Alkenyl, -C ２－２０ Alkynyl, -C(O)C １－２０ Alkyl, -C(O)C ２－２０ Alkenyl, -C(O)C ２－２０ Alkynyl, -C(O)cycloalkyl, -C(O)C １－１０ Alkylcycloalkyl; -C(O)C ２－１０ alkenylcycloalkyl, —C(O)C ２－１０ Alkynylcycloalkyl, -C(O)aryl, -C(O)C １－１０ Alkylaryl, -C(O)C ２－１０ Alkenylaryl, —C(O)C ２－１０ Alkynylaryl, —C(O)C １－１０ Alkyl C(O)R １１ , -C(O)C ２－１０ Alkenyl C(O)R １１ , -C(O)C ２－１０ AlkynylC(O)R １１ , -C(O)C １－１０ Alkyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ Alkenyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ AlkynylCH(OR １１ ) (OR １１ ), -C(O)C １－１０ Alkyl SR １１ , -C(O)C ２－１０ Alkenyl SR １１ , -C(O)C ２－１０ AlkynylSR １１ , -C(O)C １－１０ AlkylC(O)OR １１ , -C(O)C ２－１０ AlkenylC(O)OR １１ , -C(O)C ２－１０ AlkynylC(O)OR １１ , -C(O)C １－１０ AlkylC(O)SR １１ , -C(O)C ２－１０ AlkenylC(O)SR １１ , -C(O)C ２－１０ AlkynylC(O)SR １１ , 【Chemistry 2】 and R １０ is -C １－６ Alkyl, -C ２－６ Alkenyl, -C ２－６ Alkynyl, -C(O)C １－６ Alkyl, -C(O)C ２－６ Alkenyl, -C(O)C ２－６ Alkynyl, -C(O)aryl, -C(O)C １－６ Alkylaryl, -C(O)C ２－６ alkenylaryl, or -C(O)C ２－６ alkynylaryl; and R １１ is hydrogen, -C １－１０ Alkyl, -C ２－１０ Alkenyl, -C ２－１０ alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) DNA damaging chemotherapeutic agents; ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; and iii) irradiation; A method selected from the group consisting of

2. The method of claim 1 , wherein the epoxytigliane compound is administered locally to the tumor.

3. The method of claim 2 , wherein the local administration is by intratumoral injection.

4. The method of any one of claims 1 to 3, wherein the second therapeutic agent is a DNA damaging chemotherapeutic agent.

5. 5. The method of claim 4, wherein the chemotherapeutic agent is selected from a platinum chemotherapeutic agent, a DNA intercalating agent, a DNA alkylating agent, an antimetabolite, or a compound that binds to GC-rich DNA.

6. The method according to any one of claims 1 to 3, wherein the second therapeutic agent is a chemotherapeutic agent that inhibits tumor-associated host-derived cells that support tumor cell proliferation and / or invasion.

7. 7. The method of claim 6, wherein the chemotherapeutic agent is a vinca alkaloid, antimetabolite, a phosphodiesterase 5 inhibitor, amiloride, a COX-2 inhibitor, or a PEG-2 inhibitor.

8. The method of any one of claims 1 to 3, wherein the second therapeutic agent is radiation.

9. In the epoxy tigliane compound of formula (I), one or more of the following items are present: i) R １ is -CH ３ is; ii) R ２ and R ３ are independently -OC(O)C １－２０ Alkyl, -OC(O)C ２－２０ Alkenyl, -OC(O)C ２－２０ Alkynyl, -OC(O)cycloalkyl, -OC(O)C １－１０ Alkylcycloalkyl; -OC(O)C ２－１０ Alkenylcycloalkyl, —OC(O)C ２－１０ Alkynylcycloalkyl, -OC(O)aryl, -OC(O)C １－１０ Alkylaryl, -OC(O)C ２－１０ Alkenylaryl, -OC(O)C ２－１０ Alkynylaryl, -OC(O)C １－１０ Alkyl C(O)R １１ , -OC(O)C ２－１０ Alkenyl C(O)R １１ , -OC(O)C ２－１０ AlkynylC(O)R １１ , -OC(O)C １－１０ Alkyl CH(OR １１ ) (OR １１ ), -OC(O)C ２－１０ Alkenyl CH(OR １１ ) (OR １１ ), -OC(O)C ２－１０ AlkynylCH(OR １１ ) (OR １１ ), -OC(O)C １－１０ Alkyl SR １１ , -OC(O)C ２－１０ Alkenyl SR １１ , -OC(O)C ２－１０ AlkynylSR １１ , -OC(O)C １－１０ AlkylC(O)OR １１ , -OC(O)C ２－１０ AlkenylC(O)OR １１ , -OC(O)C ２－１０ AlkynylC(O)OR １１ , -OC(O)C １－１０ AlkylC(O)SR １１ , -OC(O)C ２－１０ AlkenylC(O)SR １１ , and -OC(O)C ２－１０ AlkynylC(O)SR １１ Selected from: iii) R ４ and R ５ are independently H and -CH ３ Selected from: iv) R ６ is hydrogen, -C(O)C １－６ Alkyl, -C(O)C ２－６ Alkenyl, -C(O)C ２－６ alkynyl or -C(O)aryl; v) R ７ is hydroxyl, -OC(O)C １－６ Alkyl, -OC(O)C ２－６ Alkenyl or -OC(O)C ２－６ alkynyl; and vi) R ８ is H or -CH ３ That is, The method according to any one of claims 1 to 8, wherein

10. R ２ and R ３ are independently -OC(O)C １－６ Alkyl, -OC(O)C ２－６ Alkenyl, -OC(O)C ２－６ Alkynyl, -OC(O)cycloalkyl, -OC(O)C １－６ Alkylcycloalkyl, -OC(O)C ２－６ Alkenylcycloal, -OC(O)C ２－６ 10. The method of claim 9, wherein the alkyl group is selected from alkynylcycloalkyl, and -OC(O)aryl.

11. R ２ and R ３ are independently -OC(O)C １－２０ Alkyl, -OC(O)C ２－２０ Alkenyl and OC(O)C ２－２０ 11. The method according to claim 9 or claim 10, wherein the alkyl group is selected from alkynyl.

12. The compound of formula (I) may be a compound of formula (II): 【Chemistry 3】 or a geometric isomer, stereoisomer, or pharma- ceutically acceptable salt thereof, In the formula, R ６ , R ７ , and R ９ is as defined in claim 1, The method according to any one of claims 1 to 11.

13. The epoxy tigliane compound of formula (I) is the following compound: 12-tigloyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12,13-di-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12-Hexanoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12,13-dihexanoyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12-Myristoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12-tigloyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13-pentahydroxy-20-acetyloxy-1-tiglien-3-one; 12-Myristoyl-13-acetyloxy-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12-propanoyl-13-(2-methylbutanoyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; 12,13-ditigloyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; and 12-(2-methylbutanoyl)-13-tigloyl-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tiglien-3-one; or a pharma- ceutically acceptable salt thereof, The method of claim 1 , wherein the compound is selected from the group consisting of

14. The method of any one of claims 1 to 13, wherein the epoxytigliane compound and the second therapeutic agent are administered separately and simultaneously.

15. The method according to any one of claims 1 to 14, wherein the tumor is a cutaneous tumor or a subcutaneous tumor.

16. The method of any one of claims 1 to 15, wherein the tumor is a squamous cell carcinoma, an adenocarcinoma or a sarcoma.

17. The method of any one of claims 1 to 16, wherein the tumor is head and neck cancer.

18. 18. The method of claim 16 or 17, wherein the tumor is a squamous cell carcinoma of the skin or mucosa.

19. The epoxy tigliane compound is (4S,5S,6R,7S,8R,9R,10S,11R,12R,13S,14R)-12-(2E)-2-methyl-2-enoethyl-13((2S)-2-methylbutyroyl)-6,7-epoxy-4,5,9,12,13,20-hexahydroxy-1-tigliane-3-one.

19. The method according to any one of claims 1 to 18.

20. 20. The method of claim 19, wherein the second therapeutic agent is selected from cisplatin, 5-fluorouracil, doxorubicin, and radiation.

21. 1. A combination of an epoxytigliane compound and a second therapeutic agent for use in the treatment of tumors, said epoxytigliane compound being a compound of formula (I): 【Chemistry 4】 or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R １ is hydrogen or C １－６ is alkyl; R ２ is -OH or -OR ９ and R ３ is -OH or -OR ９ and R ４ and R ５ are independently hydrogen and C １－６ alkyl; R ６ is hydrogen or -R １０ and R ７ is hydroxy or OR １０ and R ８ is hydrogen or C １－６ is alkyl; R ９ is -C １－２０ Alkyl, -C ２－２０ Alkenyl, -C ２－２０ Alkynyl, -C(O)C １－２０ Alkyl, -C(O)C ２－２０ Alkenyl, -C(O)C ２－２０ Alkynyl, -C(O)cycloalkyl, -C(O)C １－１０ Alkylcycloalkyl; -C(O)C ２－１０ alkenylcycloalkyl, —C(O)C ２－１０ Alkynylcycloalkyl, -C(O)aryl, -C(O)C １－１０ Alkylaryl, -C(O)C ２－１０ Alkenylaryl, —C(O)C ２－１０ Alkynylaryl, —C(O)C １－１０ Alkyl C(O)R １１ , -C(O)C ２－１０ Alkenyl C(O)R １１ , -C(O)C ２－１０ AlkynylC(O)R １１ , -C(O)C １－１０ Alkyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ Alkenyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ AlkynylCH(OR １１ ) (OR １１ ), -C(O)C １－１０ Alkyl SR １１ , -C(O)C ２－１０ Alkenyl SR １１ , -C(O)C ２－１０ AlkynylSR １１ , -C(O)C １－１０ AlkylC(O)OR １１ , -C(O)C ２－１０ AlkenylC(O)OR １１ , -C(O)C ２－１０ AlkynylC(O)OR １１ , -C(O)C １－１０ AlkylC(O)SR １１ , -C(O)C ２－１０ AlkenylC(O)SR １１ , -C(O)C ２－１０ AlkynylC(O)SR １１ , 【Chemistry 5】 and R １０ is -C １－６ Alkyl, -C ２－６ Alkenyl, -C ２－６ Alkynyl, -C(O)C １－６ Alkyl, -C(O)C ２－６ Alkenyl, -C(O)C ２－６ Alkynyl, -C(O)aryl, -C(O)C １－６ Alkylaryl, -C(O)C ２－６ alkenylaryl, or -C(O)C ２－６ alkynylaryl; and R １１ is hydrogen, -C １－１０ Alkyl, -C ２－１０ Alkenyl, -C ２－１０ alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) DNA damaging chemotherapeutic agents; ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; and iii) irradiation; A combination selected from.

22. 1. Use of an epoxytigliane compound in the manufacture of a first medicament, and a second chemotherapeutic agent in the manufacture of a second medicament, as a combination therapy for treating a tumor, wherein the epoxytigliane compound is a compound of formula (I): 【Chemistry 6】 or a geometric or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R １ is hydrogen or C １－６ is alkyl; R ２ is -OH or -OR ９ and R ３ is -OH or -OR ９ and R ４ and R ５ are independently hydrogen and C １－６ alkyl; R ６ is hydrogen or -R １０ and R ７ is hydroxy or OR １０ and R ８ is hydrogen or C １－６ is alkyl; R ９ is -C １－２０ Alkyl, -C ２－２０ Alkenyl, -C ２－２０ Alkynyl, -C(O)C １－２０ Alkyl, -C(O)C ２－２０ Alkenyl, -C(O)C ２－２０ Alkynyl, -C(O)cycloalkyl, -C(O)C １－１０ Alkylcycloalkyl; -C(O)C ２－１０ alkenylcycloalkyl, —C(O)C ２－１０ Alkynylcycloalkyl, -C(O)aryl, -C(O)C １－１０ Alkylaryl, -C(O)C ２－１０ Alkenylaryl, —C(O)C ２－１０ Alkynylaryl, —C(O)C １－１０ Alkyl C(O)R １１ , -C(O)C ２－１０ Alkenyl C(O)R １１ , -C(O)C ２－１０ AlkynylC(O)R １１ , -C(O)C １－１０ Alkyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ Alkenyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ AlkynylCH(OR １１ ) (OR １１ ), -C(O)C １－１０ Alkyl SR １１ , -C(O)C ２－１０ Alkenyl SR １１ , -C(O)C ２－１０ AlkynylSR １１ , -C(O)C １－１０ AlkylC(O)OR １１ , -C(O)C ２－１０ AlkenylC(O)OR １１ , -C(O)C ２－１０ AlkynylC(O)OR １１ , -C(O)C １－１０ AlkylC(O)SR １１ , -C(O)C ２－１０ AlkenylC(O)SR １１ , -C(O)C ２－１０ AlkynylC(O)SR １１ , 【Chemistry 7】 and R １０ is -C １－６ Alkyl, -C ２－６ Alkenyl, -C ２－６ Alkynyl, -C(O)C １－６ Alkyl, -C(O)C ２－６ Alkenyl, -C(O)C ２－６ Alkynyl, -C(O)aryl, -C(O)C １－６ Alkylaryl, -C(O)C ２－６ alkenylaryl, or -C(O)C ２－６ alkynylaryl; and R １１ is hydrogen, -C １－１０ Alkyl, -C ２－１０ Alkenyl, -C ２－１０ alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted; and The second therapeutic agent is: i) a DNA damaging chemotherapeutic agent; and ii) chemotherapeutic agents that inhibit tumor-associated host-derived cells that support tumor cell proliferation and / or invasion; Selected from, use.

23. 1. Use of an epoxytigliane compound in the manufacture of a medicament in combination with radiation therapy for treating a tumor; said epoxytigliane compound being a compound of formula (I): 【Chemistry 8】 or a geometric isomer or stereoisomer or a pharma- ceutically acceptable salt thereof; During the ceremony, R １ is hydrogen or C １－６ is alkyl; R ２ is -OH or -OR ９ and R ３ is -OH or -OR ９ and R ４ and R ５ are independently hydrogen and C １－６ alkyl; R ６ is hydrogen or -R １０ and R ７ is hydroxy or OR １０ and R ８ is hydrogen or C １－６ is alkyl; R ９ is -C １－２０ Alkyl, -C ２－２０ Alkenyl, -C ２－２０ Alkynyl, -C(O)C １－２０ Alkyl, -C(O)C ２－２０ Alkenyl, -C(O)C ２－２０ Alkynyl, -C(O)cycloalkyl, -C(O)C １－１０ Alkylcycloalkyl; -C(O)C ２－１０ alkenylcycloalkyl, —C(O)C ２－１０ Alkynylcycloalkyl, -C(O)aryl, -C(O)C １－１０ Alkylaryl, -C(O)C ２－１０ Alkenylaryl, —C(O)C ２－１０ Alkynylaryl, —C(O)C １－１０ Alkyl C(O)R １１ , -C(O)C ２－１０ Alkenyl C(O)R １１ , -C(O)C ２－１０ AlkynylC(O)R １１ , -C(O)C １－１０ Alkyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ Alkenyl CH(OR １１ ) (OR １１ ), -C(O)C ２－１０ AlkynylCH(OR １１ ) (OR １１ ), -C(O)C １－１０ Alkyl SR １１ , -C(O)C ２－１０ Alkenyl SR １１ , -C(O)C ２－１０ AlkynylSR １１ , -C(O)C １－１０ AlkylC(O)OR １１ , -C(O)C ２－１０ AlkenylC(O)OR １１ , -C(O)C ２－１０ AlkynylC(O)OR １１ , -C(O)C １－１０ AlkylC(O)SR １１ , -C(O)C ２－１０ AlkenylC(O)SR １１ , -C(O)C ２－１０ AlkynylC(O)SR １１ , 【Chemistry 9】 and R １０ is -C １－６ Alkyl, -C ２－６ Alkenyl, -C ２－６ Alkynyl, -C(O)C １－６ Alkyl, -C(O)C ２－６ Alkenyl, -C(O)C ２－６ Alkynyl, -C(O)aryl, -C(O)C １－６ Alkylaryl, -C(O)C ２－６ alkenylaryl, or -C(O)C ２－６ alkynylaryl; and R １１ is hydrogen, -C １－１０ Alkyl, -C ２－１０ Alkenyl, -C ２－１０ alkynyl, cycloalkyl or aryl; wherein each alkyl, alkenyl, alkynyl, cycloalkyl or aryl group is optionally substituted.

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