Pharmaceutical combinations for treatment of cancer

Abstract

The name of the invention is a pharmaceutical combination for the treatment of cancer. A pharmaceutical combination comprising (A): a polyunsaturated fatty acid and (B): a chemotherapeutic compound for simultaneous, separate or sequential use in the treatment of cancer in a human patient.

Description

[0001] This application is a divisional application. The application date of the original application is May 15, 2018, the application number is (201880032629.0), and the name is “Drug combination for treating cancer”. Technical Field

[0002] The present invention relates to a pharmaceutical combination comprising (A) a polyunsaturated fatty acid and (B) a chemotherapeutic compound for simultaneous, separate or sequential use in treating cancer in a human patient. Background Art

[0003] EP2409963B1 (Lipopharma - filed in 2010) describes the use of 1,2-derivatives of polyunsaturated fatty acids (known as D-PUFA) compounds in the treatment of cancer.

[0004] The fatty acid derivative compound has the formula:

[0005] COOR 1 -CHR 2 -(CH 2 )a-(CH=CH-CH 2 )b-(CH 2 )c-CH 3

[0006] An example of a preferred compound is:

[0007] COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (182A1)

[0008] The above compound (182A1) is further described in detail in the article "Erazo et al.; Clinical Cancer Research; 22(10), May 15, 2016" - this compound is referred to as "ABTL0812" in this article, and this term is used in this article.

[0009] The article describes that ABTL0812 induces autophagy-mediated cancer cell death without activating apoptosis. The article states:

[0010] [p2515]:

[0011] “Most current anticancer treatments activate apoptosis, and resistance to chemotherapy is a major challenge in cancer (24). Autophagy-mediated cell death has emerged as an alternative approach to kill cancer cells without inducing resistance to apoptosis-inducing drugs (25).”

[0012] [p2517]:

[0013] “On the other hand, mTORCI activation is often associated with resistance to anti-tumor drugs (6). Since ABTL0812 is a potent inhibitor of the Akt / mTORC1 axis, its combination with standard chemotherapeutic drugs could prove effective in treating drug-resistant or apoptosis-refractory tumors.”

[0014] On the filing date of this application, the applicant's website (AbilityPharma- www.abilitypharma.com ) including the news section (all inventors of the present application have transferred all rights related to this document to the applicant of the present application and the priority application EP17382282.6 of May 16, 2017 - in other words, the web page publication content of the present applicant discussed below can be regarded as the so-called "disclosure content derived from the inventor" - (that is, the disclosed subject matter must be attributed to the inventor, one or more co-inventors, or another person who obtained the subject matter directly or indirectly from the inventor or co-inventors).

[0015] The news on November 22, 2016 reads:

[0016] "Ability Pharmaceuticals, SL, a Catalan biopharmaceutical company, announced today the opening of its first Phase 2 clinical trial with its novel targeted anticancer agent ABTL0812 to evaluate its efficacy and safety as first-line treatment in combination with paclitaxel and carboplatin in 80 patients with advanced or recurrent endometrial cancer or squamous cell carcinoma of the lung (…)

[0017] In preclinical cancer models, ABTL0812 was effective as a single agent with an excellent safety profile across multiple cancer types (lung cancer, endometrial cancer, pancreatic cancer, and neuroblastoma). The compound also synergized with chemotherapeutic agents (taxanes, platinum compounds, and gemcitabine) in these models without increasing their toxicity.”

[0018] The news on December 14, 2016 reads:

[0019] “In preclinical studies, ABTL0812 has demonstrated efficacy in pancreatic cancer as a single agent and in combination with taxanes, platinum compounds, and gemcitabine, showing synergistic effects (8- to 90-fold) inducing tumor regression without increasing the toxicity associated with chemotherapy (…)

[0020] ABTL0812 is currently in Phase 2 as a first-line treatment in combination with chemotherapeutic agents in patients with endometrial cancer or squamous cell lung cancer.”

[0021] Regarding the combined use of ABTL0812 compound with other chemotherapeutic agents, neither the above-mentioned Erazo article nor the news published by the applicant (AbilityPharma) discloses any valid experimental data, i.e. Statements related to combinations can be are considered merely statements without any valid verifiable experimental data to support them. .

[0022] The published news refers to Phase II studies - as is known in the art, the fact that Phase II studies are ongoing means that Phase I studies have already been completed and based on this information the skilled person can only conclude that the results regarding safety and tolerability in humans and the pharmacokinetic studies are positive - i.e. this does not provide information about the possible therapeutic effect in human patients and in particular does not provide information about any possible synergistic effects of the combination. The skilled person will only know the results of the evaluation of whether the drug is therapeutically effective after the completion of the Phase II trial - at the date of filing of this application, no experimental data relevant to this article from the Phase II trial has been published. Summary of the invention

[0023] The problem underlying the present invention is to provide an improved cancer treatment.

[0024] As discussed above, the compound COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 Referred to herein as ABTL0812.

[0025] The working examples herein provide a number of detailed experimental data demonstrating the effects of the above-discussed combination of the ABTL0812 compound with other chemotherapeutic agents. Significant synergy .

[0026] As discussed in further detail herein - the experimental data provided herein are based on established in vitro and in vivo (e.g., mouse) experimental tests - therefore, based on the experimental data provided herein, it is credible / reliable that the synergistic effects associated with the present invention can be obtained in human cancer patients.

[0027] Example 4 of this article discusses HumanPreliminary results obtained from the pivotal trial - These results show that the synergistic effect associated with the use of the ABTL0812 compound in combination with paclitaxel and carboplatin in patients with advanced endometrial cancer or squamous cell carcinoma also exists in humans, and in this sense the obtained human clinical trials are positive.

[0028] Based on the knowledge of the prior art, the skilled artisan could not have foreseen with a reasonable expectation of success the remarkable synergistic effects described experimentally herein.

[0029] As discussed above with respect to the web page disclosures of the present applicant (AbilityPharma), the statements relating to the combination in these web page disclosures can be regarded as merely statements without any valid verifiable experimental data to support them - obviously, based on these web page disclosures, it is credible / reliable that the synergistic effects related to this article can be obtained in human cancer patients.

[0030] In short, the examples in this paper illustrate the following aspects in particular:

[0031] Example 1: - In vitro experiment:

[0032] 1.1: ABTL0812 and docetaxel have in vitro synergistic effects in representative non-small cell lung adenocarcinoma cell lines, where ABTL0812 increases the IC 50 It was reduced by more than 80-fold—i.e., it greatly increased docetaxel cytotoxicity;

[0033] 1.2: ABTL0812 and paclitaxel have in vitro synergistic effects in 4 different lung cancer cell lines, where ABTL0812 increases the IC of paclitaxel depending on the cell line. 50 It was reduced by 2 to 7 times, which means that the cytotoxicity of paclitaxel was increased;

[0034] 1:3: ABTL0812 and gemcitabine have in vitro synergistic effects in representative pancreatic cancer cell lines, where ABTL0812 increases the IC of gemcitabine 50 It was reduced by 7-fold—i.e., it greatly increased gemcitabine cytotoxicity;

[0035] 1:4: ABTL0812 and carboplatin have in vitro synergistic effects in representative endometrial cancer cell lines, where ABTL0812 increases the IC of carboplatin 50 It was reduced by 3-fold—that is, it increased carboplatin cytotoxicity;

[0036] 1:5: ABTL0812 and retinoic acid have in vitro synergistic effects in representative neuroblastoma cancer cell lines;

[0037] 1:6: ABTL0812 and paclitaxel have in vitro synergistic effects in representative breast cancer cell lines, in which ABTL0812 IC of paclitaxel 50 Reduced by 3 times ——That is, the cytotoxicity of paclitaxel was increased.

[0038] Example 2: - In vivo experiments:

[0039] 2.1: In a representative in vivo mouse model, low-dose ABTL0812 enhanced the lung cancer antitumor activity of docetaxel without negative toxic effects.

[0040] 2.2: In a representative in vivo mouse model, the combination of ABTL0812 + paclitaxel and carboplatin (P / C) treatment showed a significant improvement in survival in a squamous cell carcinoma (SCC) model, with a survival of 75% at 20 days after treatment, compared to 0% in the ABTL0812 and vehicle groups and 25% in the P / C group.

[0041] 2.3: In a representative in vivo mouse model, the combination of ABTL0812+P / C exhibited synergistic effects (relative to the effects of each drug alone in relation to adenocarcinoma lung cancer) as significant tumor volume reductions were observed in animals treated with the combination versus control and chemotherapeutic-treated animals;

[0042] 2.4: In a representative in vivo mouse model, the combination of ABTL0812 + pemetrexed and cisplatin demonstrated synergistic effects (relative to the effects of the chemotherapeutic agents alone in relation to adenocarcinoma lung cancer) as significant tumor volume reductions were observed in animals treated with the combination versus control and chemotherapeutic-treated animals;

[0043] 2.5: In a representative in vivo mouse model, the combination of ABTL0812 + paclitaxel demonstrated synergistic effects (relative to the effects of each drug alone in relation to endometrial cancer) as significant tumor volume reduction was observed in animals treated with the combination versus control animals;

[0044] 2.6: In a representative in vivo mouse model from patient-derived xenografts, the combination of ABTL0812+P / C showed synergistic effects related to endometrial cancer, showing significantly higher tumor volume reduction compared to P / C, which also showed significant tumor volume reduction compared to the vehicle group during the first 47 days;

[0045] 2.7: In a representative in vivo mouse model, the combination of ABTL0812 + paclitaxel / gemcitabine showed synergistic effects related to pancreatic cancer, showing significantly higher tumor volume reduction compared to P / Gm alone. P / Gm also showed higher tumor volume reduction compared to vehicle;

[0046] 2.8: In a representative in vivo mouse model, the combination of ABTL0812 + Nab-paclitaxel / gemcitabine showed synergistic effects related to pancreatic cancer, showing significantly higher tumor volume reduction compared to Nab-P / Gm alone. Nab-P / Gm also showed higher tumor volume reduction compared to the vehicle;

[0047] 2.9: In a representative in vivo mouse model, the combination of ABTL0812 + cisplatin showed synergistic effects related to neuroblastoma cancer, where the combination of ABTL0812 and cisplatin stabilized tumor progression for a longer period of time;

[0048] 2.10: In a representative in vivo mouse model, the combination of ABTL0812+doxorubicin showed synergistic effects related to breast cancer, where the combination of ABTL0812 and paclitaxel showed a higher tumor volume reduction compared to vehicle.

[0049] The experimental data presented herein are based on established in vitro and in vivo (eg, mouse) experimental tests - therefore, based on the experimental data presented herein, it is credible / reliable that the synergistic effects associated with this article can be obtained in human cancer patients.

[0050] Examples of chemotherapeutic agents tested in the working examples herein include:

[0051] Taxanes: paclitaxel (Taxol), Nab-paclitaxel (albumin-bound paclitaxel), and docetaxel;

[0052] Platinum-based agents: carboplatin and cisplatin;

[0053] Nucleotide analogs and prodrug analogs: gemcitabine;

[0054] Folic acid antimetabolites: pemetrexed;

[0055] Anthracyclines: doxorubicin;

[0056] Thus, different groups / classes of chemotherapeutic agents have been tested and all showed significant synergistic effects when used in combination with the ABTL0812 compound.

[0057] Based on the experimental data presented herein, it is credible that the positive synergistic effects described herein can be obtained with most relevant chemotherapeutic agents.

[0058] As discussed above, most current anticancer treatments activate apoptosis, and all of the other chemotherapeutic agents tested above activate apoptosis.

[0059] ABTL0812 induces autophagy-mediated cancer cell death without activating apoptosis. The experimental data presented herein indicate that ABTL0812 combined with chemotherapeutic agents can unexpectedly increase the level of apoptosis, but this is not the underlying mechanism of ABTL0812.

[0060] The experimental data presented herein indicate that chemotherapeutic agents (e.g., docetaxel) generally have the following characteristics:

[0061] An amount of docetaxel that produces 100% therapeutic effect => causes toxicity Y.

[0062] An amount of docetaxel that produces a 50% therapeutic effect => results in reduced toxicity.

[0063] Docetaxel in an amount that produces 50% therapeutic effect + ABTL0812 in an amount that produces 50% therapeutic effect => results in 100% effect, and toxicity is maintained at the same reduced level.

[0064] Surprisingly for the present inventors, the effect of chemotherapeutic agents (eg, docetaxel) can be significantly enhanced by combination with ABTL0812 without significantly increasing toxicity.

[0065] The ABTL0812 compound is similar in structure and function to other 1,2-derivatives of polyunsaturated fatty acids (D-PUFA) compounds described in EP2409963B1 discussed above.

[0066] Thus, it would appear that essentially all of the fatty acid derivative compounds of EP 2409963 B1 would have a synergistic effect relevant herein in combination with a chemotherapeutic agent.

[0067] Therefore, a first aspect of the present invention relates to a pharmaceutical combination comprising:

[0068] (A): as formula COOR 1 -CHR 2 -(CH 2 )a-(CH=CHCH 2 )b-(CH 2 )c-CH 3 A compound of polyunsaturated fatty acids, a pharmaceutically acceptable salt thereof or a combination thereof, wherein

[0069] (i) a can be any integer value between 0 and 7,

[0070] (ii) b can be any integer value between 2 and 7,

[0071] (iii) c can be any integer value between 0 and 7,

[0072] (iv) R1 It is H, Na, K, CH 3 , CH 3 -CH 2 or PO(O-CH 2 -CH 3 ) 2 ,and

[0073] (v)R 2 It is OH, OCH 3 、O-CH 3 COOH, CH 3 , Cl, CH 2 OH, OPO(O-CH 2 -CH 3 ) 2 , NOH, F, HCOO or N(OCH 2 CH 3 ) 2 ;

[0074] and

[0075] (B): Chemotherapeutic compound

[0076] They are used for simultaneous, separate or sequential use in the treatment of cancer in human patients.

[0077] As will be appreciated by those skilled in the art, the chemotherapeutic agent of compound (B) of the first aspect is certainly not a compound within the scope of compound (A) of the first aspect.

[0078] As will be appreciated by those skilled in the art, it is not necessary for the combination therapy discussed herein to be administered simultaneously, e.g., as a single composition, or sequentially, e.g., as two different compositions, of the two compounds (A) and (B). Importantly, when the second compound / agent is administered, an effective amount of the first administered compound / agent is already present in the patient's body.

[0079] Thus, the term "combination" of the first aspect relates herein to various combinations of compounds (A) and (B), such as in the form of a single pharmaceutical composition, in the form of a combined mixture of different pharmaceutical formulations / compositions of a plurality of single active compounds (e.g., a "tank mix"), and in the combined use of a plurality of single active ingredients when applied in a sequential manner (i.e., one after the other, separated by a reasonably short period of time (e.g., a few hours or days)) or simultaneously. The order in which compounds (A) and (B) are applied is not essential.

[0080] The combination of compounds (A) and (B) can be formulated for simultaneous, separate or sequential administration. In particular, if administration is not simultaneous, the compounds are administered relatively close to each other. In addition, the compounds are administered in the same or different dosage forms or by the same or different routes of administration, for example, one compound can be administered topically and the other compound can be administered orally. The combination of the two compounds can be administered, for example, in the following form:

[0081] - in the form of a combination as part of the same pharmaceutical formulation, the two compounds are always administered simultaneously;

[0082] - in the form of a combination of two units / compositions, each unit / composition having one of the substances enabling simultaneous, sequential or separate administration;

[0083] For example, compound (A) is administered separately from compound (B) (ie in two units) but at the same time.

[0084] In another suitable example, compound (A) is administered first, followed by compound (B) either separately or sequentially - alternatively, compound (B) is administered first, followed by compound (A) either separately or sequentially.

[0085] The term "drug" (e.g., with respect to a "pharmaceutical composition") is to be understood in accordance with the art - that is, it refers to a preparation / composition that is in a form that permits the biological activity of the active ingredient to be effective and that is physiologically tolerable (i.e., does not contain other components that are unacceptably toxic to the subject to which the composition will be administered). In particular, the term "pharmaceutically acceptable" means that it is approved by a state or federal regulatory agency or included in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals (particularly humans).

[0086] Embodiments of the invention are described below by way of example only.

[0087] A combination of one preferred embodiment described herein with another preferred embodiment described herein is a more preferred embodiment. BRIEF DESCRIPTION OF THE DRAWINGS

[0088] Figure 1 ： ABTL0812 showed in vitro synergy with docetaxel in the A549 human lung adenocarcinoma cell line. The cytotoxicity of ABTL0812, docetaxel, and the combination of both drugs was significantly reduced due to the addition of low concentrations (approximately its IC 50 half) of the ABTL0812, its IC 50 The results show that the cytotoxicity of docetaxel is enhanced by 86 times. The results show the average of two independent experiments. For further details, see the working examples of this article.

[0089] Figure 2 ： ABTL0812 showed in vitro synergy with paclitaxel in A549 and H1975 human lung adenocarcinoma cell lines and in H157 and HTB182 human lung squamous cell carcinoma cell lines. Cytotoxicity of ABTL0812, paclitaxel, and the combination of both drugs in all four different lung cancer cell lines. 50 half) of the ABTL0812, its IC 50 Depending on the cell line, the reduction was 2 to 7 times, so an enhancement of paclitaxel cytotoxicity was observed in all four cell lines. The results show the average of two independent experiments for each cell line. For further details, see the working examples of this article.

[0090] Figure 3 ： ABTL0812 showed in vitro synergy with gemcitabine in the MiaPaca2 human pancreatic cancer cell line. Cytotoxicity of ABTL0812, gemcitabine, and the combination of both drugs. 50 half of the ABTL0812, its IC 50 The cytotoxicity of gemcitabine was reduced by 7-fold, so an enhancement of gemcitabine cytotoxicity can be observed in the presence of ABTL0812. The results show the average of two independent experiments. For further details, see the working examples herein.

[0091] Figure 4 ： ABTL0812 showed in vitro synergy with carboplatin in the Ishikawa human endometrial cancer cell line. The cytotoxicity of ABTL0812, carboplatin, and the combination of both drugs was significantly reduced due to the addition of low concentrations (approximately its IC 50 half of the ABTL0812, its IC 50 The results are shown in Table 1. The results are shown in Table 1. The results are shown in Table 1. The results are shown in Table 1. The results are shown in Table 1. The results are shown in Table 1.

[0092] Figure 5 ： ABTL0812 showed in vitro synergy with retinoic acid in LAI-5S and SK-N-BE(2) human neuroblastoma cell lines. Cytotoxicity of ABTL0812, retinoic acid, and the combination of both drugs was significantly reduced due to the presence of RA in the presence of low concentrations (approximately its IC 30When incubated with ABTL0812 (half of 100 μg / mL), the cell viability of retinoic acid (RA) in LAI-5S cells decreased from 82.8% to 23.8%, while the cell viability of RA in SK-N-BE (2) decreased from 70.4% to 34.7%, so that enhancement of RA cytotoxicity can be observed in both cell lines. The results show the average of two independent experiments. For further details, please see the working examples of this article.

[0093] Figure 6 ： ABTL0812 showed in vitro synergy with paclitaxel in the MDA-DB-231 human triple-negative breast cancer cell line. The cytotoxicity of ABTL0812, paclitaxel, and the combination of the two drugs was significantly reduced due to the addition of low concentrations (below its IC 50 ) of ABTL0812, its IC 50 The cytotoxicity of paclitaxel was reduced by 2.7 times, so the enhancement of paclitaxel cytotoxicity can be observed. The results show the average of two independent experiments. For further details, see the working examples of this article.

[0094] Figure 7 ： ABTL0812 showed an enhancement of docetaxel therapeutic effects in the A549 human lung adenocarcinoma xenograft model in vivo without increasing toxicity. Left: The anti-tumor effect of the combination of ABTL0812 and docetaxel in the a549 human lung adenocarcinoma xenograft model, showing a significant reduction in tumor growth compared to the ABTL0812, docetaxel and vehicle groups. The p value was measured between ABTL0812+docetaxel and docetaxel and between the vehicle. Right: The total weight of the different treatment groups during the entire experiment. For further details, see the working examples of this article.

[0095] Figure 8 ： ABTL0812 showed an enhancement of the therapeutic effect of paclitaxel / carboplatin (P / C) in the H157 human lung squamous cell carcinoma xenograft model in vivo without increasing toxicity. Kaplan-Meier graphs of H157 xenografts treated with ABTL0812, P / C, ABTL0812+P / C and vehicle, wherein ABTL0812+P / C showed the highest survival rate. For further details, see the working examples of this article.

[0096] Fig. 9 ：ABTL0812 showed an enhancement of the therapeutic effect of P / C without increasing toxicity in the H1975 human lung adenocarcinoma xenograft model in vivo. Left: The anti-tumor effect of the combination of ABTL0812 and P / C in the H1975 human lung adenocarcinoma xenograft model, showing the highest tumor volume reduction compared to P / C, ABTL0812 or vehicle groups. The p value is measured between ABTL0812+P / C and P / C and between ABTL0812+P / C and vehicle. Right: Total weight of different treatment groups during the entire experiment. For further details, please see the working examples herein.

[0097] Fig.10 ： ABTL0812 showed an enhancement of the therapeutic effect of pemetrexed and cisplatin in the A549 human lung adenocarcinoma xenograft model in vivo without increasing toxicity. Left: The anti-tumor effect of the combination of ABTL0812 with pemetrexed and cisplatin, showing a significant reduction in tumor growth compared to the pemetrexed and cisplatin and vehicle groups. The p value is measured between ABTL0812+pemetrexed and cisplatin and vehicle. Right: The total weight of the different treatment groups during the entire experiment. For further details, see the working examples of this article.

[0098] Fig.11 ： ABTL0812 showed an enhancement of the therapeutic effect of paclitaxel without increasing toxicity in the orthotopically implanted in vivo Ishikawa human endometrial cancer xenograft model. Left: Animals were sacrificed after three weeks of treatment, tumors were excised, and tumor volumes were determined. A significant statistical reduction was observed in animals receiving the combination of ABTL0812 + paclitaxel compared to control animals receiving only vehicle. The p value was measured between ABTL0812 + paclitaxel and vehicle. Right: Total body weight of different treatment groups throughout the experiment. For further details, see the working examples herein.

[0099] Fig.12 ： ABTL0812 showed an enhancement of the therapeutic effect of P / C in human endometrial cancer patient-derived xenografts in vivo without increasing toxicity. Left: A tumor was implanted into nude mice, which was surgically removed from a patient with serous histology, grade IIIC2, 100% myometrial infiltration, pelvic and aortic lymph nodes and lymphovascular space infiltration, and carrying p53 and PI3KCA gene mutations. The combination of ABTL0812 and P / C showed a significant reduction in tumors compared to P / C, ABTL0812 and vehicle. The p value of the figure is indicated in the figure. Right: The total weight of the different treatment groups during the entire experiment is shown below. For further details, see the working examples of this article.

[0100] Fig.13 ：ABTL0812 showed enhancement of gemcitabine and paclitaxel (Gm / P) in the in vivo MiaPAca2 human pancreatic cancer xenograft model without increasing toxicity. Left: Antitumor effect of the combination of ABTL0812+Gem / P, showing the highest tumor volume reduction compared to Gm / P, ABTL0812 or vehicle. The p value was measured between ABTL0812+gemcitabine and paclitaxel and gemcitabine and paclitaxel, between gemcitabine and between vehicle. Right: Total weight of different treatment groups during the entire experiment. For further details, see the working examples of this article.

[0101] Fig.14 ： ABTL0812 showed enhancement of gemcitabine (Gm) alone and enhancement of gemcitabine and Nab-paclitaxel (Gm / Nab-P) without increased toxicity in two in vivo MiaPAca2 human pancreatic cancer xenograft models. Left: Anti-tumor effect of the combination of ABTL0812+Gm / Nab-P, showing the highest tumor volume reduction compared to Gm / Nab-P, ABTL0812 or vehicle. Right: Anti-tumor effect of the combination of ABTL0812+Gm, showing the highest tumor volume reduction compared to Gm, ABTL0812 or vehicle. The p value is measured between ABTL0812+Gm / Nab-P and ABTL0812+Gm and vehicle. The total weight of different treatment groups during the entire experiment is as follows. For further details, please refer to the working examples of this article.

[0102] Fig.15 ： ABTL0812 shows an enhancement of the therapeutic effect of cisplatin in the in vivo SH-SY5Y human neuroblastoma xenograft model without increasing toxicity. Left: The anti-tumor effect of the combination of ABTL0812+ cisplatin, showing a reduction in the highest tumor volume compared to ABTL0812, cisplatin or vehicle. On the 10th day, the animals in the control group must be killed, and at the same time, half of the animals in the treatment group are also killed, and the remaining animals are studied for a longer time. Left: Tumor weight when killed after 10 days of treatment (by t test, p < 0.05). For further details, please refer to the working examples of this article.

[0103] Fig.16 ： ABTL0812 showed enhancement of doxorubicin therapeutic effect without increased toxicity in the in vivo MDA-DB-231 human triple-negative breast cancer xenograft model. Left: Anti-tumor effect of the combination of ABTL0812+doxorubicin, showing the highest tumor volume reduction compared to doxorubicin, ABTL0812 or vehicle. P value is measured between ABTL0812+doxorubicin and vehicle. Right: Total body weight of different treatment groups during the entire experiment. For further details, see the working examples herein.

[0104] Fig.17 ： ABTL0812 showed an enhancement of the therapeutic effect of gemcitabine and cisplatin (Gm / Cis) in the in vivo EGI-1 human cholangiocarcinoma xenograft model without increasing toxicity. Left: Anti-tumor effect of the combination of ABTL0812+Gm / Cis, showing the highest tumor volume reduction compared to Gm / Cis, ABTL0812 or vehicle. The p value is measured between ABTL0812+Gm / Cis and vehicle. Right: Total body weight of different treatment groups during the entire experiment. For further details, see the working examples herein. DETAILED DESCRIPTION

[0105] The compound (A) of the first aspect

[0106] A preferred embodiment is one in which

[0107] (i) a can be any integer value between 5 and 7,

[0108] (ii) b can be any integer value between 2 and 4,

[0109] (iii) c can be any integer value between 1 and 5.

[0110] Preferably, R 1 Can be H, Na, K, CH 3 , CH 3 -CH 2 or PO(O-CH 2 -CH 3 ) 2 ,

[0111] Preferably, R 2 Can be OH, OCH 3 、O-CH 3 COOH, CH 3 , Cl, CH 2 OH, OPO(O-CH 2 -CH 3 ) 2 , NOH, F, HCOO or N(OCH 2 CH 3 ) 2 .

[0112] In a preferred embodiment, R 1 is H, and R 2 It's OH.

[0113] In another preferred embodiment, R 1 is Na, and R 2 It's OH.

[0114] Preferably, compound (A) is at least one compound selected from the group consisting of:

[0115] COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812),

[0116] COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 3 -CH 3 (183A1),

[0117] COOH-CHOH-(CH 2 ) 3 -(CH=CH-CH 2 ) 3 -(CH 2 ) 3 -CH 3 (183A2),

[0118] COOH-CHOH-(CH 2 ) 2 -(CH=CH-CH 2 ) 4 -(CH 2 ) 3 -CH 3 (204A1),

[0119] COOH-CHOH-(CH 2 ) 2 -(CH=CH-CH 2 ) 5 -CH 3 (205A1) and

[0120] COOH-CHOH-CH 2 -(CH=CH-CH 2 ) 6 -CH 3 (226A1).

[0121] Most preferably, compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2-(CH 2 ) 3 -CH 3 (ABTL0812).

[0122] The pharmaceutically acceptable salt of compound (A) refers to any pharmaceutically acceptable salt of compound (A). As known in the art, there are a variety of known pharmaceutically acceptable salts. Examples of pharmaceutically acceptable salts include, but are not limited to, sodium (Na), potassium, acetate, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, acetate, propionate, decanoate, octanoate, acrylate, formate, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyrate, succinate ... In one embodiment, the salt of compound (A) is a sodium salt.

[0123] As will be appreciated by those skilled in the art, when referring to a preferred formula of compound (A) (e.g., ABTL0812) herein, it should be understood herein that the formula also includes its salt forms, for example, when referring to compound (A) herein as COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812), it also refers to the salts of ABTL0812.

[0124] Preferably, compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 The sodium salt of (ABTL0812).

[0125] The compound (B) of the first aspect

[0126] Preferably, compound (B) is at least one chemotherapeutic compound selected from the group consisting of:

[0127] Bifunctional alkylating agents (preferably cyclophosphamide, dichloromethyl diethylamine, chlorambucil or melphalan);

[0128] A monofunctional alkylating agent (preferably dacarbazine (DTIC), nitrosourea or temozolomide);

[0129] Anthracycline (preferably daunomycin, doxorubicin, epirubicin, idarubicin, mitoxantrone or valrubicin);

[0130] Taxane (preferably paclitaxel, docetaxel, Nab-paclitaxel or taxotere);

[0131] Epothilone (preferably patupirone, sagopilone or ixabepilone);

[0132] Deacetylase inhibitors (preferably vorinostat or romidepsin);

[0133] Topoisomerase I inhibitors (preferably irinotecan or topotecan);

[0134] Topoisomerase II inhibitors (preferably etoposide, teniposide or tafluposide);

[0135] Kinase inhibitors (preferably bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, or vismodegib);

[0136] Nucleotide analogs and / or precursor analogs (preferably azacitidine, azathioprine, capecitabine, cytarabine, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate or thioguanine);

[0137] Peptide antibiotics (preferably bleomycin or actinomycin);

[0138] Platinum-based agents (preferably carboplatin, cisplatin or oxaliplatin);

[0139] A retinoid (preferably tretinoin, alitretinoin, or bexarotene); and

[0140] Vinca alkaloids and derivatives (preferably vinblastine, vincristine, vindesine or vinorelbine).

[0141] As is understood in the art, relative to any preferred listed examples of compound (B), it is most preferred that compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH2 ) 3 -CH 3 (ABTL0812).

[0142] More preferably, compound (B) is at least one chemotherapeutic compound selected from the group consisting of:

[0143] Cyclophosphamide;

[0144] Melphalan;

[0145] Docetaxel;

[0146] Paclitaxel;

[0147] Nab-paclitaxel;

[0148] Carboplatin;

[0149] Cisplatin;

[0150] Oxaliplatin;

[0151] Methotrexate

[0152] Pemetrexed;

[0153] Azathioprine;

[0154] Capecitabine;

[0155] Fluorouracil;

[0156] Mercaptopurine;

[0157] Gemcitabine;

[0158] Bleomycin;

[0159] Actinomycin;

[0160] vincristine;

[0161] Vinblastine;

[0162] Vinorelbine;

[0163] Retinoic acid;

[0164] Temozolomide;

[0165] Daunocin

[0166] Doxorubicin;

[0167] irinotecan; and

[0168] Topotecan.

[0169] Even more preferably, compound (B) is at least one chemotherapeutic compound selected from the group consisting of:

[0170] Docetaxel;

[0171] Paclitaxel;

[0172] Nab-paclitaxel;

[0173] Carboplatin;

[0174] Cisplatin;

[0175] Oxaliplatin;

[0176] Methotrexate

[0177] Pemetrexed;

[0178] Gemcitabine;

[0179] Bleomycin

[0180] Retinoic acid;

[0181] Temozolomide;

[0182] Doxorubicin;

[0183] irinotecan; and

[0184] Topotecan.

[0185] It may be preferred that compound (B) of the first aspect comprises two or more different chemotherapeutic agents (particularly when compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812)) - For example, preferably, the compound (B) of the first aspect comprises:

[0186] paclitaxel and carboplatin;

[0187] paclitaxel and gemcitabine;

[0188] Nab-paclitaxel and gemcitabine;

[0189] gemcitabine and cisplatin;

[0190] Pemetrexed and cisplatin.

[0191] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is docetaxel - particularly wherein the cancer is lung cancer (preferably non-small cell lung adenocarcinoma). (See Examples 1.1 and 2.1 herein for examples of this preferred embodiment).

[0192] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is paclitaxel - particularly where the cancer is lung cancer (non-small cell lung cancer). (See Example 1.2 herein for an example of this preferred embodiment).

[0193] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is gemcitabine - particularly wherein the cancer is pancreatic cancer. (For examples of this preferred embodiment, see, for example, Example 1.3 herein).

[0194] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is carboplatin - particularly wherein the cancer is endometrial carcinoma. (See Example 1.4 herein for an example of this preferred embodiment).

[0195] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is retinoic acid - particularly wherein the cancer is neuroblastoma. (See Example 1.5 herein for an example of this preferred embodiment).

[0196] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is paclitaxel - particularly wherein the cancer is breast cancer (preferably triple negative breast cancer). (For an example of this preferred embodiment, see Example 1.6 herein).

[0197] Particularly preferably, compound (A) is ABTL0812 and compound (B) is paclitaxel and carboplatin - particularly where the cancer is squamous cell carcinoma (preferably non-small cell lung squamous cell carcinoma). (For an example of this preferred embodiment, see Example 2.2 herein).

[0198] Particularly preferred is that compound (A) is ABTL0812 and compound (B) is paclitaxel and carboplatin - particularly where the cancer is non-small cell lung adenocarcinoma. (See Example 2.3 herein for an example of this preferred embodiment).

[0199] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is pemetrexed and cisplatin - particularly where the cancer is non-small cell lung adenocarcinoma. (See Example 2.4 herein for an example of this preferred embodiment).

[0200] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is paclitaxel - particularly wherein the cancer is endometrial cancer. (See Example 2.5 herein for an example of this preferred embodiment).

[0201] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is paclitaxel and carboplatin - particularly wherein the cancer is endometrial cancer. (See Example 2.6 herein for an example of this preferred embodiment).

[0202] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is paclitaxel and gemcitabine - particularly wherein the cancer is pancreatic cancer. (See Example 2.7 herein for an example of this preferred embodiment).

[0203] Particularly preferably, compound (A) is ABTL0812 and compound (B) is gemcitabine, or Nab-paclitaxel and gemcitabine - particularly wherein the cancer is pancreatic cancer. (See Example 2.8 herein for an example of this preferred embodiment).

[0204] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is cisplatin - particularly wherein the cancer is neuroblastoma cancer. (See Example 2.9 herein for an example of this preferred embodiment).

[0205] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is doxorubicin - particularly where the cancer is triple negative breast cancer. (See Example 2.10 herein for an example of this preferred embodiment).

[0206] It is particularly preferred that compound (A) is ABTL0812 and compound (B) is gemcitabine and cisplatin - particularly wherein the cancer is cholangiocarcinoma. (See Example 2.11 herein for an example of this preferred embodiment).

[0207] Preferably, compound (A) is ABTL0812 and compound (B) is temozolomide and irinotecan - particularly wherein the cancer is neuroblastoma cancer.

[0208] Preferably, compound (A) is ABTL0812 and compound (B) is doxorubicin and topotecan - particularly wherein the cancer is neuroblastoma cancer.

[0209] Preferably, the pharmaceutical combination discussed herein is one in which compound (A) is ABTL0812, and in which:

[0210] - Compound (B) is docetaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma;

[0211] - Compound (B) is paclitaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma;

[0212] - Compound (B) is gemcitabine, and the cancer is pancreatic cancer;

[0213] - Compound (B) is carboplatin, and the cancer is endometrial cancer;

[0214] - Compound (B) is retinoic acid, and the cancer is neuroblastoma;

[0215] - Compound (B) is paclitaxel, and the cancer is breast cancer, preferably triple-negative breast cancer;

[0216] - Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma, preferably non-small cell lung squamous cell carcinoma;

[0217] - Compound (B) is paclitaxel and carboplatin, and the cancer is non-small cell lung adenocarcinoma;

[0218] - Compound (B) is pemetrexed and cisplatin, and the cancer is non-small cell lung adenocarcinoma;

[0219] - Compound (B) is paclitaxel, and the cancer is endometrial cancer;

[0220] - Compound (B) is paclitaxel and carboplatin, and the cancer is endometrial cancer;

[0221] - Compound (B) is paclitaxel and gemcitabine, and the cancer is pancreatic cancer;

[0222] - Compound (B) is gemcitabine, and the cancer is pancreatic cancer;

[0223] - Compound (B) is gemcitabine and Nab-paclitaxel, and the cancer is pancreatic cancer;

[0224] - Compound (B) is cisplatin and gemcitabine, and the cancer is neuroblastoma;

[0225] - Compound (B) is doxorubicin acid, and the cancer is breast cancer, preferably triple-negative breast cancer;

[0226] - Compound (B) is cisplatin and gemcitabine, and the cancer is cholangiocarcinoma;

[0227] - Compound (B) is gemcitabine, and the cancer is cholangiocarcinoma;

[0228] - compound (B) is topotecan, and the cancer is neuroblastoma;

[0229] - Compound (B) is irinotecan and the cancer is neuroblastoma; or

[0230] - Compound (B) is temozolomide and the cancer is neuroblastoma.

[0231] As discussed above, Example 4 herein discusses preliminary results obtained from human pivotal trials - these results indicate that there is also a synergistic effect associated with the use of the ABTL0812 compound in combination with paclitaxel and carboplatin in patients with advanced endometrial cancer or squamous cell carcinoma in humans, and in this sense, the obtained human clinical trials are positive.

[0232] Therefore, it is preferred that the pharmaceutical combination discussed herein is one in which compound (A) is ABTL0812, and in which:

[0233] - Compound (B) is paclitaxel and carboplatin, and the cancer is advanced endometrial cancer, preferably advanced endometrial cancer; or

[0234] - Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma.

[0235] Preferably, ABTL0812 is administered orally - preferably, the administered dose of ABTL0812 is a dose of 1200 mg to 1400 mg.

[0236] More preferably, ABTL0812 is administered orally at a starting dose of 1200 mg to 1400 mg three times a day in combination with chemotherapy.

[0237] cancer

[0238] Preferably, the cancer is at least one cancer selected from the group consisting of:

[0239] Lung cancer;

[0240] Non-small cell lung cancer;

[0241] Small cell lung cancer;

[0242] Squamous cell carcinoma;

[0243] Adenocarcinoma;

[0244] Endometrial cancer;

[0245] Pancreatic cancer;

[0246] Glioblastoma;

[0247] Breast cancer;

[0248] Head and neck cancer;

[0249] neuroblastoma; and

[0250] Biliary duct cancer.

[0251] More preferably, the cancer is at least one cancer selected from the group consisting of:

[0252] Non-small cell lung cancer;

[0253] Squamous cell carcinoma;

[0254] Endometrial cancer;

[0255] Pancreatic cancer;

[0256] Glioblastoma;

[0257] Breast cancer;

[0258] neuroblastoma; and

[0259] Biliary duct cancer.

[0260] Administration of compound (A) and / or compound (B):

[0261] As discussed above, it is not necessary for the combination therapy discussed herein to be administered simultaneously, e.g., as a single composition, or sequentially, e.g., as two different compositions, of the two compounds (A) and (B). Importantly, when the second compound / agent is administered, an effective amount of the first administered compound / agent is already present in the patient's body.

[0262] It may be preferred that the pharmaceutical combinations discussed herein are a single composition comprising both Compound (A) and Compound (B).

[0263] Compound (A) (particularly ABTL0812) is preferably administered orally.

[0264] The administration dose of compound (A) (particularly ABTL0812) is preferably a dose of 200 mg to 6000 mg (preferably 2000 mg), more preferably a dose of 300 mg to 1600 mg, and even more preferably a dose of 450 mg to 1450 mg.

[0265] More preferably, the administration dose of compound (A) (particularly ABTL0812) is preferably a total dose of 200 mg to 6000 mg (preferably 2000 mg) per day, more preferably a total dose of 300 mg to 1600 mg per day, and even more preferably a total dose of 450 mg to 1450 mg per day. Preferably, the total dose is provided by administration 1 to 5 times per day, more preferably 2 to 4 times per day, and most preferably 3 times per day.

[0266] Thus, if the total dose is, for example, 1200 mg per day and is provided by 3 administrations per day - then each of the 3 administrations may, for example, be 400 mg.

[0267] With respect to Compound (B), the preferred route of administration will generally depend on the chemotherapeutic agent of interest.

[0268] The preferred administration routes of the preferred compound (B) are briefly described as follows:

[0269] Docetaxel; - preferably administered intravenously via an infusion solution

[0270] Paclitaxel; - preferably administered intravenously via an infusion solution

[0271] Carboplatin; - preferably administered intravenously via an infusion solution

[0272] Cisplatin; - preferably administered intravenously via an infusion solution

[0273] Gemcitabine; - preferably administered intravenously via an infusion solution

[0274] Nab-paclitaxel - Preferably administered intravenously via an infusion suspension

[0275] Pemetrexed; - preferably administered intravenously via an infusion solution

[0276] Doxorubicin; preferably administered intravenously

[0277] Aspects / embodiments of the invention in the so-called claim format:

[0278] 1. A drug combination comprising:

[0279] (A): as the formula COOR 1 -CHR 2 -(CH 2 )a-(CH=CHCH 2 )b-(CH 2 )c-CH 3 A compound of polyunsaturated fatty acids, a pharmaceutically acceptable salt thereof or a combination thereof, wherein

[0280] (i) a can be any integer value between 0 and 7,

[0281] (ii) b can be any integer value between 2 and 7,

[0282] (iii) c can be any integer value between 0 and 7,

[0283] (iv) R 1 It is H, Na, K, CH 3 , CH 3 -CH 2 or PO(O-CH 2 -CH 3 ) 2 ,and

[0284] (v)R 2 It is OH, OCH 3 、O-CH 3 COOH, CH 3 , Cl, CH 2 OH, OPO(O-CH 2 -CH 3 ) 2 , NOH, F, HCOO or N(OCH 2 CH 3 ) 2 ;

[0285] and

[0286] (B): Chemotherapeutic compound

[0287] They are used for simultaneous, separate or sequential use in the treatment of cancer in human patients.

[0288] 2. The pharmaceutical combination according to claim 1, wherein

[0289] (i) a can be any integer value between 5 and 7,

[0290] (ii) b can be any integer value between 2 and 4, and

[0291] (iii) c can be any integer value between 1 and 5.

[0292] 3. A pharmaceutical combination according to any one of the preceding claims, wherein R 1 is H, and R 2 It's OH.

[0293] 4. The pharmaceutical combination according to any one of the preceding claims, wherein compound (A) is at least one compound selected from the group consisting of:

[0294] COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812),

[0295] COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 3 -CH 3 (183A1),

[0296] COOH-CHOH-(CH 2 ) 3 -(CH=CH-CH 2 ) 3 -(CH 2 ) 3 -CH 3 (183A2),

[0297] COOH-CHOH-(CH 2 ) 2 -(CH=CH-CH 2 ) 4 -(CH 2 ) 3 -CH 3 (204A1),

[0298] COOH-CHOH-(CH 2 ) 2 -(CH=CH-CH 2 ) 5 -CH 3 (205A1) and

[0299] COOH-CHOH-CH 2 -(CH=CH-CH 2 ) 6 -CH 3 (226A1).

[0300] 5. A pharmaceutical combination according to any one of the preceding claims, wherein compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812) or a pharmaceutically acceptable salt thereof.

[0301] 6. The pharmaceutical combination according to claim 5, wherein compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 The sodium salt of (ABTL0812).

[0302] 7. The pharmaceutical combination according to any one of the preceding claims, wherein the cancer is at least one cancer selected from the group consisting of:

[0303] Lung cancer;

[0304] Non-small cell lung cancer;

[0305] Squamous cell carcinoma;

[0306] Adenocarcinoma;

[0307] Endometrial cancer;

[0308] Serous cancer;

[0309] Pancreatic cancer;

[0310] Glioblastoma cancer;

[0311] Drug-resistant recurrent breast cancer;

[0312] Head and neck cancer;

[0313] Neuroblastoma and

[0314] Biliary duct cancer.

[0315] 8. The pharmaceutical combination according to any one of the preceding claims, wherein compound (B) is at least one chemotherapeutic compound selected from the group consisting of:

[0316] Docetaxel;

[0317] Paclitaxel;

[0318] Carboplatin;

[0319] Cisplatin;

[0320] Gemcitabine;

[0321] Nab-paclitaxel;

[0322] Retinoic acid;

[0323] Temozolomide;

[0324] Irinotecan;

[0325] Doxorubicin; and

[0326] Topotecan.

[0327] 9. The pharmaceutical combination according to claim 8, wherein compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 )3 -CH 3 (ABTL0812) or a pharmaceutically acceptable salt thereof.

[0328] 10. The pharmaceutical combination according to claim 9, wherein

[0329] - Compound (B) is docetaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma;

[0330] - Compound (B) is paclitaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma;

[0331] - Compound (B) is gemcitabine, and the cancer is pancreatic cancer;

[0332] - Compound (B) is carboplatin, and the cancer is endometrial carcinoma;

[0333] - Compound (B) is retinoic acid, and the cancer is neuroblastoma cancer;

[0334] - Compound (B) is paclitaxel acid, and the cancer is breast cancer, preferably triple-negative breast cancer;

[0335] - Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma, preferably non-small cell lung squamous cell carcinoma;

[0336] - Compound (B) is paclitaxel and carboplatin, and the cancer is non-small cell lung adenocarcinoma;

[0337] - Compound (B) is pemetrexed and cisplatin, and the cancer is non-small cell lung adenocarcinoma;

[0338] - Compound (B) is paclitaxel, and the cancer is endometrial cancer;

[0339] - Compound (B) is paclitaxel and carboplatin, and the cancer is endometrial cancer;

[0340] - Compound (B) is paclitaxel and gemcitabine, and the cancer is pancreatic cancer;

[0341] - Compound (B) is gemcitabine, and the cancer is pancreatic cancer;

[0342] - Compound (B) is gemcitabine and Nab-paclitaxel, and the cancer is pancreatic cancer;

[0343] - Compound (B) is cisplatin and gemcitabine, and the cancer is neuroblastoma cancer;

[0344] - Compound (B) is doxorubicin acid, and the cancer is breast cancer, preferably triple-negative breast cancer; or

[0345] - Compound (B) is cisplatin and gemcitabine, and the cancer is cholangiocarcinoma.

[0346] 11. The pharmaceutical combination according to any one of the preceding claims, wherein the pharmaceutical combination is a single composition comprising both Compound (A) and Compound (B).

[0347] 12. The pharmaceutical combination according to any one of the preceding claims, wherein compound (A) is administered orally.

[0348] 13. The pharmaceutical combination according to any one of the preceding claims, wherein compound (A) is administered in a total dose of 200 mg to 2000 mg per day.

[0349] 14. The pharmaceutical combination according to any one of claims 12 to 13, wherein compound (A) is COOH-CHOH-(CH 2 ) 6 -(CH=CH-CH 2 ) 2 -(CH 2 ) 3 -CH 3 (ABTL0812) or a pharmaceutically acceptable salt thereof.

[0350] 15. The pharmaceutical combination according to claim 14, wherein

[0351] - Compound (B) is docetaxel and is administered intravenously via an infusion solution;

[0352] - Compound (B) is paclitaxel and is administered intravenously via an infusion solution;

[0353] - Compound (B) is carboplatin and is administered intravenously via an infusion solution;

[0354] - Compound (B) is cisplatin and it is administered intravenously via an infusion solution;

[0355] - Compound (B) is gemcitabine and is administered intravenously via an infusion solution;

[0356] - Compound (B) is Nab-paclitaxel and is administered intravenously via an infusion suspension;

[0357] - Compound (B) is temozolomide and it is administered orally (e.g. in capsule form);

[0358] - Compound (B) is irinotecan and is administered intravenously via an infusion solution;

[0359] - Compound (B) is doxorubicin and it is administered intravenously; or

[0360] - Compound (B) is topotecan and it is administered intravenously via an infusion solution.

[0361] Examples

[0362] Example 1 : Combination of ABTL0812 with different chemotherapeutic agents - in vitro assays 1.1：

[0364] Cell viability assay of ABTL0812 combined with docetaxel in lung cancer

[0365] Research reference: ABT-EI-

[0366] Research Location: Protein Kinase and Cell Signaling Group, UAB

[0367] GLP Compliance: No

[0368] Test compound: ABTL0812 (Batch 006 / 2010)

[0369] Reference compound: Docetaxel (Fluka, 01855-5MG-F, batch 1425738V)

[0370] Test system: A549 (human lung cancer) cell line

[0371] Objective: To evaluate the effect of the combination of ABTL0812 and docetaxel on cell viability. Docetaxel is a cytotoxic compound used in lung cancer; therefore, potential treatment combinations for lung cancer may involve the use of ABTL0812 and docetaxel.

[0372] Methods: A549 cells were treated with increasing concentrations of ABTL0812 (3-300 μM), docetaxel (0.01-100 μM), or a combination of both (IC 50 Herein, a fixed concentration of 20 μM ABTL0812 and 0.01-100 μM docetaxel) were incubated together for three days (FBS 0.5%). Cell viability was assessed by MTT assay in all cases, and the IC values ​​of ABTL0812, docetaxel and the combination were calculated. 50. Finally, the combination index (CI) for assessing synergy was calculated according to the Chou and Talalay method (Chou 2006; Chou 2010) as follows: CI = (D)1 / (Dx)1+(D)2 / (Dx)2, where CI <1, =1 and >1 indicate synergy, additivity and antagonism, respectively. In the denominator, (Dx)1 means that D1 "alone" inhibits the system by x%, while (Dx)2 means that D2 "alone" inhibits the system by x%. In the numerator, the "combination" of (D)1 and (D)2 also inhibits by x%. The results shown are the average of two independent experiments.

[0373] Results: As expected, ABTL0812 and docetaxel were cytotoxic when used independently. Low concentrations of ABTL0812 (20 μM, equivalent to its IC 10 ) greatly increased docetaxel cytotoxicity. The IC of docetaxel in the presence of ABTL0812 50 The decrease was >80-fold, from 1.7 μM to 0.02 μM (see table below and in this article). Figure 1 ).

[0374]

[0375] Then, the potential synergy of ABTL0812 and docetaxel was calculated according to the method of Chou and Talalay (Chou 2006; Chou 2010). The combination of the two drugs was synergistic across the entire range of activity, with a CI = 0.47 at 50% cell viability. This CI indicates synergy.

[0376] Conclusion: ABTL0812 and docetaxel have in vitro synergistic effects in lung adenocarcinoma cell line A549. Suboptimal concentration of ABTL0812 (20 μM) reduces the IC 50 The decrease was more than 80-fold. These results provide an opportunity for in vivo combination of the two drugs in lung cancer. Docetaxel is a first-line drug for multiple stages of lung cancer; therefore, combination with ABTL0812 has a potential beneficial effect, as synergy between the two drugs has been observed to increase their cytotoxic effects in lung cancer cells. 1.2：

[0378] Cell viability assay of ABTL0812 combined with paclitaxel in lung cancer

[0379] Research reference: LN3-T30 and ABT-EI-048

[0380] Research Location: Protein Kinase and Cell Signaling Group, UAB

[0381] GLP Compliance: No

[0382] Test compound: ABTL0812 (Batch 002 / 2012)

[0383] Reference compound: Paclitaxel (SelleckChem, S1150-10MG, Batch 09)

[0384] Test systems: A549 (human lung cancer) cell line with mutant KRAS; H157 (human non-small cell lung squamous cell carcinoma with mutant PTEN; HTB182: human non-small cell lung squamous cell carcinoma and H1975: human non-small cell lung adenocarcinoma with mutant PI3KCA

[0385] Objective: To evaluate the effect of the combination of ABTL0812 and paclitaxel on cell viability. Paclitaxel is a cytotoxic compound used in lung cancer; therefore, potential therapeutic combinations for lung cancer may involve the use of ABTL0812 and paclitaxel.

[0386] Methods: A549 cells were treated with increasing concentrations of ABTL0812 (3-300 μM), paclitaxel (0.003-1 μM), or a combination of both (using IC 50 The following fixed concentrations of ABTL0812 and 0.003-1 μM paclitaxel) were incubated together in DMEM containing 0.1% FBS for 72 hours. H157, H1957 and HTB-812 cells were incubated with increasing concentrations of paclitaxel (0.001-10 μM) (alone) and paclitaxel with IC 50 The following combinations of fixed concentrations of ABTL0812 were incubated together in DMEM containing 0.1% FBS for 48 h. Cell viability was assessed by MTT assay in all cases, and the IC of paclitaxel and the combination was calculated. 50 The concentrations of the different cell line combinations are:

[0387] A549 cells: ABTL0812 IC 50 =49 μM. Paclitaxel combined with 10 (IC10), 20 (IC25) and 30 μM (IC35) ABTL0812

[0388] H157 cells: ABTL0812 IC 50 = 23 μM. Paclitaxel combined with 10 (IC20) and 15 (IC35) μM ABTL0812

[0389] H1957 cells: ABTL0812 IC50 = 43 μM. Paclitaxel combined with 10 (IC10) and 20 (IC20) μM ABTL0812

[0390] HTB-812 cells: ABTL0812 IC50 = 29 μM. Paclitaxel was combined with 10 (IC20) and 15 (IC35) μM ABTL0812.

[0391] Finally, as mentioned in the previous section, for paclitaxel IC 50 The combination index (CI) was calculated.

[0392] Results: ABTL0812 and paclitaxel were cytotoxic in all four lung cancer cell lines. When used independently, the addition of 15, 20, or 30 μM ABTL0812 increased paclitaxel cytotoxicity in the A549 lung cancer cell line. The combined IC 50 Lower than the IC of each individual drug 50 , and the IC of paclitaxel was observed 50 The paclitaxel IC 50 The reduction in paclitaxel was synergistic, with CIs of 0.34, 0.28, and 0.22 for 15, 20, and 30 μM ABTL0812, respectively. In H157 cells, the synergy observed was not as strong as that observed in A549 cells, which only enhanced paclitaxel cytotoxicity when combined with 15 μM ABTL0812 (IC35), increasing the paclitaxel IC 50 The decrease from 4.19 to 2.39 μM was 1.75-fold, showing synergy with a CI of 0.7. In the case of H1957 cells, when ABTL0812 was added at 10 μM (IC10), there was no synergy, and the IC 50 The value did not change (3.68 vs. 3.47 μM), but when ABTL0812 was added at 20 μM (IC20), the IC 50 The IC value of paclitaxel in H1957 decreased by 6.5 times from 3.68 to 0.56 μM. 50 The reduction was synergistic with a CI of 0.3. Finally, HTB-812 cells showed good synergy at both tested concentrations, with a 3-fold decrease from 2.71 to 0.81 μM when ABTL0812 was added at 10 μM (IC20) and a 3.6-fold decrease from 2.71 to 0.75 μM when ABTL0812 was added at 20 μM (IC35). These paclitaxel ICs in HTB-812 cells 50 The reductions were synergistic, with CIs of 0.4 and 0.5, respectively. For further details, see Figure 2 .

[0393] Conclusions: ABTL0812 and paclitaxel have in vitro synergistic effects in all four lung cancer cell lines tested, regardless of their mutational status. Suboptimal concentrations of ABTL0812 reduce the IC 50 These results provide an opportunity for in vivo combination of the two drugs in lung cancer. 1.3：

[0395] Cell viability assay of ABTL0812 alone or in combination with gemcitabine in pancreatic cancer

[0396] Research reference: LN1-T56-T58

[0397] Research Location: Protein Kinase and Cell Signaling Group, UAB

[0398] GLP Compliance: No

[0399] Test compound: ABTL0812 (Batch 006 / 2010)

[0400] Reference compound: Gemcitabine (Sigma, G6423, batch 041M4727V)

[0401] Test system: MiaPaca2 (human pancreatic cancer)

[0402] Objective: To investigate the potential synergy when adding ABTL0812 to gemcitabine in the pancreatic cancer cell line MiaPaca2. Gemcitabine is considered the standard of care for the treatment of most types of pancreatic cancer. In many cases (mostly for advanced pancreatic cancer), it is administered in combination with other drugs (Ghaneh and Neoptolemos 2007); therefore, it would be interesting to know if there is any additive effect between the two drugs.

[0403] Methods: MiaPaca2 cells were treated with gemcitabine (0.01-100 μM), ABTL0812 (3-300 μM), or a combination of both (IC 50 Below, a fixed concentration of 25 μM ABTL0812 was seeded in a 24-well plate with 0.01-100 μM gemcitabine) and placed in an incubator for 72 hours (0.5% FBS). Cell viability was studied by MTT assay, and several parameters were determined to evaluate possible synergy. First, the IC of each individual drug or combination was calculated. 50 The synergy (CI) was then calculated as described above.

[0404] Results: Combined IC 50 Lower than the IC of each individual drug 50 , because the IC of gemcitabine was observed 50Note that the ABTL0812 concentration selected for the combination experiment has very low activity alone (<10% cytotoxicity), but it enhances the cytotoxicity of gemcitabine - see the table below and in this article Figure 3 .

[0405]

[0406] For non-constant combination ratios, the potential synergy of ABTL0812 with gemcitabine was calculated according to the method of Chou and Talalay (Chou 2006; Chou 2010). The combination of the two drugs was synergistic across the entire range of activity, with a CI = 0.65 at 50% cell viability. This CI indicates synergy.

[0407] Conclusion: ABTL0812 and gemcitabine have in vitro synergistic effects in the pancreatic cancer cell line MiaPaca2. Suboptimal concentration of ABTL0812 (25 μM) reduces the IC of gemcitabine. 50 These results provide an opportunity for in vivo combination of the two drugs. 1.4：

[0409] Cell viability assay study of ABTL0812 alone or in combination with carboplatin in endometrial cancer Reference: ABT-EI-065

[0410] Research Location: Protein Kinase and Cell Signaling Group, UAB

[0411] GLP Compliance: No

[0412] Test compound: ABTL0812 (Batch 006 / 2010)

[0413] Reference compound: Carboplatin (Sigma, C2538)

[0414] Test system: Ishikawa (human endometrial carcinoma)

[0415] Objective: To investigate the potential synergy when adding ABTL0812 to carboplatin in the endometrial cancer cell line Ishikawa. Carboplatin is considered the standard of care for the treatment of most types of endometrial cancer. Therefore, it would be interesting to know if there is any additive effect between the two drugs.

[0416] Methods: Ishikawa cells were seeded in 24-well plates with increasing concentrations of carboplatin (1-300 μM) in the presence of 4 μM ABTL0812 (equivalent to IC10) for 48 h (0.5% FBS). Cell viability was studied by MTT assay, and several parameters were determined to evaluate possible synergy. First, the IC of each individual drug or combination was calculated. 50 .

[0417] Results: Combined IC 50 Lower than the IC of each individual drug 50 , because the IC of carboplatin was observed 50 Note that the ABTL0812 concentration chosen for the combination experiment had very low activity alone (<10% cytotoxicity), but it enhanced the cytotoxicity of gemcitabine - see Figure 4 .

[0418] Conclusion: ABTL0812 and carboplatin have an in vitro synergistic effect in the endometrial cancer cell line Ishikawa. Suboptimal concentration of ABTL0812 (4 μM) reduces the IC of carboplatin. 50 These results provide an opportunity for in vivo combination of the two drugs. 1.5：

[0420] Cell viability assay study of ABTL0812 alone or in combination with retinoic acid in neuroblastoma Reference: ABT-EI-055

[0421] Study Location: Pediatric Cancer Translational Research Laboratory (VHIR)

[0422] GLP Compliance: No

[0423] Test compound: ABTL0812 (Batch 006 / 2010)

[0424] Reference compound: Retinoic acid (Sigma, R2625)

[0425] Test systems: SK-N-BE(2): human neuroblastoma cell line and LA1-5S: a clonal subline of the neuroblastoma cell line LA-N-1

[0426] Objective: To evaluate the effect of ABTL0812 in combination with retinoic acid (RA) on cell viability in neuroblastoma cell lines SK-N-BE(2) and LA1-5S.

[0427] Methods: LA1-5S and SK-N-BE(2) cells were incubated with IC 50The following fixed concentrations of ABTL0812 (30 μM), increasing concentrations of 10 μM, 20 μM and 30 μM retinoic acid or a combination of the two were incubated together. 10 μM retinoic acid is the pharmacological dose administered orally to neuroblastoma patients in a Phase I clinical trial (Villablanca et al., 1995). Cells were treated for 24 hours in IMDM containing 0.5% FBS. In all cases, cell viability was assessed by crystal violet assay. Different doses were evaluated in six parallel samples, and the results shown are the average of two independent experiments. GraphPad Statistical analysis was performed using the 5.0 software according to the T test principle (*p<0.05; **p<0.01; ***p<0.001).

[0428] Results: ABTL0812 exhibited mild cytotoxicity in both LA1-5S and SK-N-BE(2) neuroblastoma cell lines when used as a single agent at low concentrations. Retinoic acid concentrations of 10 μM and 20 μM were even less effective. The combination of 30 μM ABTL0812 and retinoic acid produced high cytotoxicity in both neuroblastoma cell lines. The percentage of dead / non-proliferating cells in any combination was higher than that of each agent alone, indicating a synergistic effect. The increase in cell death was statistically significant at all concentrations (***p<0.001). For further details, see the article. Figure 5 .

[0429] Conclusion: The combination of ABTL0812 and RA has a strong synergistic effect, which enhances their in vitro cytotoxic activity in neuroblastoma cell lines SK-N-BE(2) and LA1-5S. RA is commonly used in the clinic to treat neuroblastoma at the minimal residual disease stage, therefore, the present data encourage further investigation of this combination for the treatment of neuroblastoma. 1.6：

[0431] Cell viability assay of ABTL0812 alone or in combination with paclitaxel in breast cancer

[0432] Research reference: To be determined

[0433] Study Location: Target Laboratory, UdG

[0434] GLP Compliance: No

[0435] Test compound: ABTL0812 (Batch 006 / 2010)

[0436] Reference compound: Paclitaxel (SelleckChem, S1150)

[0437] Test system: MDB-DA-231 (human triple-negative breast cancer)

[0438] Objective: To investigate the potential synergy when adding ABTL0812 to paclitaxel in the triple-negative breast cancer cell line MDB-DA-231.

[0439] Methods: MDB-DA-231 cells were seeded in 24-well plates with increasing concentrations of paclitaxel (1-100 nM) in the presence of 5, 10, and 20 μM ABTL0812 (all doses were below the IC25) for 48 h (0.5% FBS). Cell viability was studied by MTT assay, and several parameters were determined to evaluate possible synergy. First, the IC25 of each drug alone or in combination was calculated. 50 .

[0440] Results: Combined IC 50 Lower than the IC of each individual drug 50 , because the IC of paclitaxel was observed 50 Note that the ABTL0812 concentration selected for the combination experiment had very low activity alone (<10% cytotoxicity), but it enhanced the cytotoxicity of paclitaxel, showing strong synergy with CI values ​​of 0.3, 0.2, and 0.16 for 5, 10, and 20 μM ABTL0812, respectively - see the table below and in the text Figure 6 .

[0441] Compound IC50 ABTL0812 29μM Paclitaxel 8,7nM Paclitaxel + 5 μM ABTL0812 5,1nM Paclitaxel + 10 μM ABTL0812 3,2nM Paclitaxel + 20 μM ABTL0812 3,2nM

[0442] Conclusion: ABTL0812 and carboplatin have an in vitro synergistic effect in the endometrial cancer cell line Ishikawa. Suboptimal concentration of ABTL0812 (4 μM) reduces the IC of carboplatin. 50 These results provide an opportunity for in vivo combination of the two drugs.

[0443] Example 2 : Combination of ABTL0812 with different chemotherapeutic agents - in vivo assay 2.1：

[0445] A549 xenografts in mice in combination with docetaxel

[0446] Research reference: ALM-IDIBAPS

[0447] Study setting: Molecular and Translational Oncology Research Group, IDIBAPS, Hospital Clinic of Barcelona.

[0448] GLP Compliance: No

[0449] Test compound: ABTL0812 (Batch 002 / 2012)

[0450] Reference compound: docetaxel.

[0451] Test system: Nu / nu male mice

[0452] Objective: To investigate the antitumor activity of ABTL0812 alone and in combination with docetaxel, a reference drug for the treatment of NSCLC.

[0453] Methods: Mice were injected with 5x10 6 A549 cells were used to induce tumor formation. After 20 days, when the tumor volume was approximately 50 mm 3 At 14:00, animals were randomized into groups and started different treatments. ABTL0812 was administered orally at 30 mg / kg / day 5 times a week. Docetaxel 5 mg / kg was administered intraperitoneally once a week (Coxon et al., 2012). Tumor volume and body weight were monitored 3 times a week.

[0454] Results: ABTL0812 significantly reduced tumor volume compared to control animals (ANOVA followed by t test). The efficacy of ABTL0812 is indeed similar to that observed with docetaxel treatment. Interestingly, ABTL0812 enhanced the anti-tumor effect of docetaxel. Statistical analysis showed that the combination treatment significantly improved the reduction of tumor growth compared to docetaxel alone (t test, p<0.001). In addition, no decrease in body weight or hematological counts was observed in any treatment group (including treatment groups where ABTL0812 was administered together with docetaxel) (not shown), indicating that the combination had no toxic effects. Regarding the anti-tumor effect of the combination of ABTL0812 and docetaxel in the A549 lung cancer xenograft model, all treatments significantly reduced tumor volume compared to the control at the time of sacrifice (*, ANOVA followed by t test analysis). In addition, the combination of 30 mg / kg ABTL0812 + docetaxel was significantly more effective than docetaxel alone (**p<0.01, t-test). On the other hand, no effect on body weight was observed for any of the individual treatments or their combination. For further details, see the Figure 7 .

[0455] Conclusion: ABTL0812 reduced tumor growth in a lung cancer xenograft model derived from A549 cells. ABTL0812 had efficacy similar to SOC docetaxel in this model. ABTL0812 and docetaxel as monotherapy similarly reduced tumor volume in a lung cancer xenograft model derived from A549 cells. ABTL0812 enhanced the antitumor activity of docetaxel without toxic effects. These results suggest that the combination therapy of ABTL0812 plus docetaxel may have clinical significance for the treatment of lung cancer. 2.2：

[0457] Efficacy of ABTL0812 in combination with paclitaxel and carboplatin in human squamous NSCLC (H157) xenografts in mice

[0458] Research reference: ALM-IDIBAPS

[0459] Study setting: Molecular and Translational Oncology Research Group, IDIBAPS, Hospital Clinic of Barcelona.

[0460] GLP Compliance: No

[0461] Test compound: ABTL0812 (Batch K102E)

[0462] Test system: Nu / nu male mice

[0463] Objective: To investigate the survival of ABTL0812 alone and in combination with paclitaxel and carboplatin (P / C) in a human squamous NSCLC xenograft model using Kaplan-Meier analysis.

[0464] Methods: Mice were injected with 5x10 6 H157 cells were added to induce tumor formation. When the tumor volume was approximately 100 mm 3 At 14:00, animals were randomly divided into groups (n=8 per group) and started different treatments. The different conditions studied were vehicle, ABTL0812 120 mg / kg orally per day, 15 mg / kg carboplatin + 50 mg / kg paclitaxel by ip route, and another group receiving a combination of these two regimens. ABTL0812 was always administered two days before and two days after the first P / C administration, maintaining 4 doses of ABTL0812 and one dose of P / C per week. Tumor volume was monitored 3 times per week. For Kaplan-Meier plotting, the endpoint criterion for excluding animals from the study was a tumor volume greater than >1000mm 3 Or different animal welfare indicators confirmed by the ethics committee. Maintain treatment in different groups until all animals in each group reach 1000mm 3or welfare-related endpoints, except for the ABTL0812+P / C group, in which mice had to 3 sacrificed before terminating the experimental procedure.

[0465] Results: In Kaplan-Meier analysis, the combination of ABTL0812 treatment and P / C showed the most effective treatment. Figure 8 As shown in, the combination of ABTL0812+P / C was the most effective treatment in terms of survival rate, showing a significant benefit over other groups. On day 20 after the start of treatment, ABTL0812+P / C treatment showed 75% survival without any relevant toxicity signs, compared to 0% in the ABTL0812 and P / C group and 20% in the vehicle group.

[0466] Conclusion: Endpoint criteria were established based on different measurements of animal welfare indicators and indicated endpoint decisions. When the health status of the animals is stable, 1000 mm 3 The tumor volume of 100 μg / ml was set as the endpoint criterion. Under these conditions, the combination of ABTL0812+P / C treatment showed a significant increase in survival measured by Kaplan-Meier analysis in the H157 squamous NSCLC xenograft model, with 75% survival 20 days after treatment, compared to 0% survival in ABTL0812 and vehicle and 25% survival in the P / C group.

[0467] 2.3: Efficacy of ABTL0812 in combination with paclitaxel and carboplatin in human adenocarcinoma NSCLC (H1975) xenografts in mice

[0468] Research reference: ABT-EI-049

[0469] Study setting: Molecular and Translational Oncology Research Group, IDIBAPS, Hospital Clinic of Barcelona.

[0470] GLP Compliance: No

[0471] Test compound: ABTL0812 (Batch 002 / 2012)

[0472] Reference compounds: Paclitaxel (Selleckchem #S1150) Carboplatin (Sigma Aldrich #C2538)

[0473] Test system: Nu / nu male mice

[0474] Objective: To investigate the antitumor activity of ABTL0812 alone and in combination with paclitaxel and carboplatin, one of the reference treatments for NSCLC, in human lung adenocarcinoma xenografts.

[0475] Methods: H1975 cell line was routinely cultured in DMEM 10% FBS, and cells in the exponential growth phase were harvested and counted for tumor inoculation. Mice were injected with 2.5x10 6 H1957 cells suspended in 50 μl of growth medium without FBS and 50 μl of Matrigel (Corning #354234). Tumor volume was monitored 3 days per week and when tumors reached 100 mm 3 (between 50 and 150mm 3 Animals were homogenously distributed into four treatment groups that showed similar mean intra-group tumor volumes (excluding tumors less than 50 mm 3 Or more than 150mm 3 of tumors to minimize differences).

[0476] The treatment groups were:

[0477] - Vehicle group (n=7): Oral treatment with 200 μl water + 5% glycerol four days a week; and two ip injections of 100 μl saline solution once a week

[0478] -ABTL0812 (n=9): Oral administration of 200 μl 120 mg / kg ABTL0812 (resuspended in water + 5% glycerol) 5 times a week

[0479] - Paclitaxel / Carboplatin (n=9): 100 μl 15 mg / kg paclitaxel treatment ip; and 100 μl 5 mg / kg carboplatin ip once a week

[0480] -ABTL0812+paclitaxel / carboplatin (n=9): 200 μl 120 mg / kg ABTL0812 treatment orally four days a week; and 100 μl 15 mg / kg paclitaxel ip and 100 μl 5 mg / kg carboplatin ip once a week

[0481] Results: The combination of ABTL0812 administered with paclitaxel and carboplatin exhibited the best in vivo antitumor effect in xenografts derived from H1957 cells. Although the administration of paclitaxel and carboplatin reduced tumor volume compared to the vehicle group, ABTL0812 administered alone exhibited a similar tumor volume reduction with an improved trend, while the triple combination of ABTL0812 + paclitaxel and carboplatin exhibited the highest tumor volume reduction with a significant difference. In addition, a slight decrease in body weight was observed during the first week of treatment in the triple combination group, while it remained stable for the rest of the experiment. (No decrease in body weight or hematological counts was observed in any treatment group (including the treatment group where ABTL0812 was administered together with docetaxel) (not shown), indicating that this combination had no toxic effects. For further details, see this article. Fig. 9 .

[0482] Conclusions: Orally administered ABTL0812 reduced tumor growth in a lung cancer xenograft model derived from H1975 cells. In this model, ABTL0812 had efficacy similar to that of SOC paclitaxel + carboplatin. In addition, ABTL0812 enhanced the antitumor activity of paclitaxel / carboplatin without toxic effects. These results suggest that combination therapy of ABTL0812 plus paclitaxel / carboplatin may have clinical significance for the treatment of lung cancer.

[0483] 2.4: Efficacy of ABTL0812 in combination with pemetrexed and cisplatin in human adenocarcinoma NSCLC (A549) xenografts in mice

[0484] Research reference: ABT-EI-052

[0485] Research Location: Protein Kinase and Cell Signaling Group, UAB

[0486] GLP Compliance: No

[0487] Test compound: ABTL0812 (Batch K102E)

[0488] Reference compounds: Pemetrexed (Sigma Aldrich #PHR1596) and Cisplatin (Sigma Aldrich #P4394)

[0489] Test system: Nu / nu male mice

[0490] Objective: The aim of this study was to evaluate the antitumor effect of ABTL0812 alone or in combination with pemetrexed and cisplatin (standard of care chemotherapy) in the treatment of a subcutaneous xenograft model of lung cancer in immunosuppressed nude mice transplanted with human lung adenocarcinoma A549 cells.

[0491] Methods: A549 cell line was routinely cultured in DMEM 10% FBS, and cells in the exponential growth phase were harvested and counted for tumor inoculation. Fifty female nude mice aged 8 weeks were injected with 5x10 6 MiaPaca2 cells suspended in 50 μl of growth medium without FBS and 50 μl of Matrigel (Corning #354234). Tumor volume was monitored 3 days per week and when tumors reached 100 mm 3 (between 50 and 150mm 3 Animals were homogenously distributed into three treatment groups that showed similar mean intra-group tumor volumes (excluding tumors less than 50 mm 3 Or more than 150mm 3 of tumors to minimize differences).

[0492] The treatment groups were:

[0493] - Vehicle group (n=7): 200 μl water + 5% glycerol treatment orally 4 times a week; and 200 μl saline buffer (chemotherapeutic vehicle) ip twice a week

[0494] - Pemetrexed / cisplatin (n=20): 100 μl 100 mg / kg pemetrexed ip twice a week; and 100 μl 2 mg / kg cisplatin ip once a week

[0495] -ABTL0812 + pemetrexed / cisplatin (n=20): 200 μl 120 mg / kg ABTL0812 (resuspended in water + 5% glycerol) orally, 4 times a week; and 100 μl 100 mg / kg pemetrexed ip, and 100 μl 2 mg / kg cisplatin ip, once a week

[0496] Results: Figure 1 As shown in, the combination of ABTL0812 administered with pemetrexed and cisplatin showed a strong anti-tumor effect, and enhanced the therapeutic effect of pemetrexed / cisplatin, thereby achieving a significant reduction in tumor volume compared to the pemetrexed / cisplatin treatment group. From the 33rd day of treatment, ABTL0812+pemetrexed / cisplatin showed a significant tumor reduction compared to pemetrexed / cisplatin, and the difference was more significant in the next few days until the last day of treatment (the 41st day). From the 22nd day of treatment until the last day of treatment (the 41st day), both treatment groups (pemetrexed / cisplatin and ABTL0812+pemetrexed / cisplatin) showed a significant tumor volume reduction compared to the vehicle group.

[0497] In terms of treatment-induced toxicity, Figure 2The time course of total body weight for all four groups monitored over the 41 days of treatment is shown. Although no signs of toxicity or clinical pathology were observed, a slight decrease was observed in the ABTL0812 + pemetrexed / cisplatin group compared to the remaining groups during the first week of treatment due to a decrease in food intake on the days when chemotherapy was administered. No additional clinical pathology signs were observed in any group. For further details, see the Fig.10 .

[0498] Conclusion: As previously reported, ABTL0812 reduced tumor growth in a lung cancer xenograft model derived from A549 cells. In this model, ABTL0812 enhanced the antitumor activity of pemetrexed and cisplatin without toxic effects. Pemetrexed and cisplatin treatment is the most common first-line treatment option for patients with lung adenocarcinoma, so these results suggest that the combination of ABTL0812 plus pemetrexed and cisplatin may be clinically meaningful for the treatment of patients with lung cancer. 2.5：

[0500] Efficacy study of the combination of ABTL0812 and paclitaxel in an endometrial in situ model in mice Study location: Xenopat (Barcelona, ​​Spain)

[0501] GLP study: No

[0502] Test compounds: ABTL0812 (Batch 001R / 2014), Paclitaxel (Teva)

[0503] Test system: Athymic nude-Foxn1nu female mice

[0504] Objective: To evaluate the antitumor efficacy of orally administered ABTL0812 in combination with ip paclitaxel in the Ishikawa orthotopic model of endometrial cancer.

[0505] Methods: A 3 mm 3 Ishikawa cell line derived tumor blocks. Before starting drug treatment, all animals were weighed and tumor volume was assessed by palpation. Based on the tumor volume of mice, mice were divided into several groups using a randomized block design. Paclitaxel was administered ip every 7 days (15 mg / kg). ABTL0812 was administered by oral gavage, and its administration schedule was 5 days of medication and 2 days of drug withdrawal (120 mg / kg / day). In general, animals were divided into four administration groups as shown in the following table.

[0506] Table. Administration groups in the orthotopic model of endometrial cancer.

[0507]

[0508] According to the formula V = π / 6xLxW 2 Tumor volume was estimated, where L is the long axis of the tumor and W is the short axis of the tumor. During routine monitoring, animals were examined for tumor growth and the effects of treatment on normal behavior, such as activity, visual estimation of food, weight gain / loss (weight was measured twice a week during drug administration), dull eyes / hair, and any other abnormal effects. Deaths and observed clinical signs were recorded based on the number of animals in each subset.

[0509] Since orthotopic tumors can only be measured at the time of sacrifice, a group of animals (n=2-3 per group) were sacrificed after one week of treatment to determine early effects on tumor growth. Most animals were sacrificed after 3 weeks of treatment (n=5-7 per group).

[0510] Results: In a group of mice (n=36), 3 mm 3 Ishikawa cell line derived tumor fragments. No effects on animal behavior were recorded throughout the experimental treatment period. Animals treated with the combination with paclitaxel showed a reduction in tumor weight gain, and some weight loss was partially recovered at the end of the treatment period. Given that this weight loss compared to the control group did not reach 10%, it was not considered toxic. Animals were sacrificed after one or three weeks of drug treatment and tumor volumes were determined as indicated in the methods. No differences were observed in those animals treated for one week. However, in those animals treated for three weeks, an additive effect was observed with respect to those animals treated with the combination of paclitaxel + ABTL0812 - see the present invention. Fig.11 .

[0511] Conclusion: The combination of ABTL0812 + paclitaxel has shown synergistic effects (relative to the effects of each drug alone) as significant tumor volume reduction was observed in animals treated with the combination compared to control animals. At the same time, some body weight loss was observed in animals treated with the combination. However, this effect was not considered toxic. 2.6：

[0513] Efficacy study of ABTL0812 in combination with paclitaxel and carboplatin in a patient-derived xenograft (PDX) endometrial model in mice

[0514] Research reference: ABT-EI-043

[0515] Study location: VHIR

[0516] GLP Compliance: No

[0517] Test compound: ABTL0812 (Batch K102E=MEI-014-15)

[0518] Reference compounds: Paclitaxel (Selleckchem #S1150) Carboplatin (Sigma Aldrich #C2538)

[0519] Test system: Athymic nude-Foxn1nu female mice

[0520] Objective: To investigate the antitumor activity of ABTL0812 alone and in combination with paclitaxel and carboplatin (P / C) in a subcutaneously transplanted PDX model in nude mice.

[0521] Methods: Several nude mice were implanted in one flank with tumors surgically removed from a patient with serous histology, grade IIIC2, 100% myometrial invasion, and invasion of pelvic and aortic lymph nodes and lymphovascular spaces, and carrying p53 and PI3KCA gene mutations. 3 Afterwards, the tumors were extracted from the mice, cut into 2 mm long pieces, and re-implanted into one side of the abdomen of 40 mice. 3 At , mice were randomly assigned to treatment groups and dosed as follows: vehicle (n=11); ABTL0812: 120 mg / kg orally, 5 times a week (n=10); paclitaxel / carboplatin (P / C): intraperitoneal P: 50 mg / kg / C: 15 mg / kg (n=12); and ABTL0812+P / C: ABTL0812 (n=10), always administered two days before and two days after the first P / C administration, maintaining 4 doses of ABTL0812 and one dose of P / C per week; the doses were the same as when given separately.

[0522] The effectiveness of the treatment was measured by the effect of the treatment on tumor growth (measured by its volume). During the study, the health status of the animals and drug-induced toxicity were determined by animal body weight. Tumor size evolution and tumor weight were evaluated by two-way ANOVA (day-by-day analysis).

[0523] To simulate a Phase II clinical trial design (in which ABTL0812 will be administered chronically after a P / C cycle), P / C treatment was removed from the P / C and ABTL0812+P / C groups, and ABTL0812 was maintained chronically. Tumor volumes were measured for an additional 17 days after removal of P / C treatment.

[0524] Results: When the tumor reached about 100 mm 3 A group of 40 mice was randomly assigned to four treatment groups. Fig.12As shown in , ABTL0812+P / C showed the highest efficacy and statistically significant tumor growth reduction compared to vehicle alone, ABTL0812 alone, and P / C alone at day 47 after the start of treatment. The improvement in treatment outcome was not associated with an increase in toxicity signs, as can be seen from the absence of significant weight loss in total body weight. ABTL0812 administered alone showed the same efficacy as P / C without any associated toxicity signs.

[0525] On day 47, P / C administration was stopped while ABTL0812 was maintained long-term (5 days per week) to determine whether maintained ABTL0812 administration could avoid or delay tumor recurrence. Although tumors continued to grow at a similar or slightly higher slope in the P / C group where mice stopped receiving treatment, the ABTL0812+P / C group where ABTL0812 administration was maintained not only showed no increase in tumor growth rate, but also showed signs of remission. The ABTL0812 group, which maintained ABTL0812 throughout the experiment, showed very similar tumor growth inhibition as P / C, maintaining a constant growth slope, indicating that there were no signs of drug resistance processes that would increase the growth rate.

[0526] Conclusion: The combination of ABTL0812+P / C has shown synergistic effects, showing significantly higher tumor volume reduction compared to P / C alone, which also showed significant tumor volume reduction during the first 47 days compared to the vehicle group. ABTL0812 administered as a monotherapy showed the same efficacy in reducing tumor growth as obtained with P / C administration. On day 15, body weight loss was only partially observed in the ABTL0812 group, which recovered in the next four days and remained stable for the rest of the experiment. None of the other groups showed any effects that were considered toxic.

[0527] To mimic a phase II clinical trial in humans, in which ABTL0812 would be administered in combination with P / C as first-line therapy and ABTL0812 would be retained long-term after the chemotherapy cycle, we removed P / C treatment while maintaining ABTL0812 at day 47. Although tumor growth was not reduced starting with P / C and ABTL0812, long-term administration of ABTL0812 effectively prevented tumor recurrence after P / C treatment in human endometrial PDXs, showing signs of remission 10 days after chemotherapy. 2.7：

[0529] Efficacy study of ABTL0812 in combination with paclitaxel / gemcitabine (P / Gm) in a human pancreatic xenograft model in mice transplanted with MiaPaca-2 cells

[0530] Research reference: ABT_EI_001_XP

[0531] Research location: Ability Virtual Lab - UAB

[0532] GLP Compliance: No

[0533] Test compound: ABTL0812 (Batch K102E=MEI-014-15)

[0534] Reference compound: Paclitaxel / Gemcitabine (P / Gm)

[0535] Test system: Athymic nude-Foxn1nu female mice

[0536] Objective: To evaluate the antitumor efficacy of ABTL0812 in combination with P / Gm by oral route in the pancreatic cancer MiaPaCa2 xenograft mouse model. P / Gm administered ip was used as positive control together with vehicle, ABTL0812 and the combination of ABTL0812+P / Gm. Efficacy was assessed by tumor growth, tolerability of the compound and toxicity indicated by the evolution of animal body weight.

[0537] Methods: Athymic female nude mice (n=9 per group) were injected subcutaneously in one flank with 0.1 ml of MiaPaca2 cells (5×10 6 cells / ml, serum-free DMEM medium, 1:1 with Matrigel). Tumor volume (TV) was measured as length x width. 2 x1 / 2, three times a week. When the average tumor volume reaches 100mm 3 At 4 pm, mice were randomly assigned to treatment groups and dosed as follows: vehicle; ABTL0812 120 mg / kg, 5 times per week; P / Gm 15 mg / kg and 60 mg / kg, twice per week; and a combination of ABTL0812+P / Gm, in which ABTL0812 was given four times per week (days 1, 2, 4, and 5), always two days prior to P / Gm administration (days 3 and 6), for a total of 4 weeks.

[0538] The effectiveness of the treatment was measured by the effect of the treatment on tumor growth (measured by its volume). During the study, the health status of the animals and the toxicity caused by the drug were determined by their body weight. The evolution of tumor size was evaluated by two-way ANOVA (day-by-day analysis) and by Student's t-test (days with significant differences).

[0539] Results: Fig.13The figure shows the effect of the combination of orally administered ABTL0812 and P / Gm on relative tumor volume and total body weight (as a measure of toxicity). The combination of ABTL0812 and P / Gm showed the highest efficacy, significantly reducing tumor volume at the indicated time compared to P / Gm alone. In addition, ABTL0812+P / Gm combination treatment showed tumor regression in all animals from the 10th day after treatment, contrary to what was observed in the remaining groups. ABTL0812 alone seemed to have a positive effect within the first 15 days after treatment, showing tumor volumes similar to those of the vehicle group in the following days. Compared with the vehicle and ABTL0812 groups, the indicated dose of P / Gm treatment reduced tumor volume, but no statistical significance was analyzed at any time. In the total body weight graph, we can see the effect of the combination of ABTL0812+P / Gm on total body weight compared to the remaining groups. Although there was a slight decrease in body weight after the first few doses of ABTL0812+P / Gm, the combination produced minimal toxicity, and there were no acute toxic effects in any animal during the experiment.

[0540] Conclusion: The combination ABTL0812+P / Gm has shown synergistic effects, with significantly higher tumor volume reduction compared to P / Gm alone. P / Gm also showed higher tumor volume reduction compared to the vehicle and ABTL0812 groups, but without statistical significance. ABTL0812 administered as a monotherapy showed the same tumor growth curve as obtained with the vehicle group. Notably, ABTL0812+P / Gm treatment could induce tumor regression in all individual animals treated with the combination, maintaining tumor volumes of approximately 100mm by day 10 of treatment 3 , and maintained at 100 mm after the 10th day of treatment 3 The following indicates tumor regression. Body weight reduction was partially observed in the ABTL0812+P / Gm group, but mice in any group did not show any effects that could be considered toxic. 2.8：

[0542] Efficacy study of the combination of ABTL0812 administered with Nab-paclitaxel / gemcitabine (Nab-Pac / Gm) in a human pancreatic xenograft model in mice transplanted with MiaPaca-2 cells

[0543] Research reference: ABT-EI-053

[0544] Research location: Ability Virtual Lab - UAB

[0545] GLP Compliance: No

[0546] Test compound: ABTL0812 (Batch K102E=MEI-014-15)

[0547] Reference compounds: Gemcitabine hydrochloride (Sigma Aldrich G6423), Nab-paclitaxel (Abraxane; ID: 3369272 Celgene)

[0548] Test system: Athymic nude-Foxn1nu female mice

[0549] Objective: The purpose of this study is to evaluate the efficacy and safety of ABTL0812 administered orally to enhance the antitumor effect of the standard of care (SOC) chemotherapy gemcitabine and the combination gemcitabine / Nab-paclitaxel administered intraperitoneally (ip) in an in vivo human pancreatic xenograft model in immunosuppressed nude mice transplanted with MiaPaca2 cells. Both chemotherapy options are the most common first-line treatments for advanced pancreatic cancer in humans.

[0550] Methods: MiaPaca2 cell line was routinely cultured and cells in exponential growth phase were harvested and counted for tumor inoculation. 55 immunodeficient athymic nude female mice were injected subcutaneously in one flank with 5x10 6 MiaPaca2 cells suspended in 50 μl of growth medium without FBS and 50 μl of Matrigel (Corning #354234). Tumor volume was monitored 3 days per week and when tumors reached 100 mm 3 (between 50 and 150mm 3 Animals were homogenously distributed into six treatment groups that showed similar mean intra-group tumor volumes and excluded tumors less than 50 mm 3 Or more than 150mm 3 of tumors to minimize differences.

[0551] The treatment groups were:

[0552] - Vehicle group (n=9): treated orally with 200 μl water + 5% glycerol 5 times a week (ABTL0812 vehicle); and ip 200 μl saline buffer (chemotherapeutic vehicle) twice a week.

[0553] - ABTL0812 (n=9): Oral treatment with 120 mg / kg (in 200 μl distilled water + 5% glycerol), 5 times a week.

[0554] - Gemcitabine (n=9): ip 60 mg / kg (in 100 μl sterile water) treatment, twice a week.

[0555] - Gemcitabine + Nab-paclitaxel (n = 8): Nab-paclitaxel was freshly prepared from powdered material (10% (m / m) in 0.9% NaCl solution) and administered ip at 5 mg / kg (in 230 μl) twice a week. Gemcitabine was administered ip at 60 mg / kg (in 100 μl sterile water) twice a week. It is important to change the injection site to avoid intestinal necrosis and keep the Nab-paclitaxel and gemcitabine injections as far away from each other as possible in the abdomen of the mouse.

[0556] - Gemcitabine + ABTL0812 (n = 10): Oral treatment with 120 mg / kg ABTL0812 (in 200 μl distilled water + 5% glycerol) 5 times a week; and ip administration of 60 mg / kg gemcitabine (in 100 μl sterile water) 2 times a week. ABTL0812 was always administered before chemotherapy in order to reduce the stress caused by ip injection, usually as the last administration.

[0557] - Gemcitabine + Nab-paclitaxel + ABTL0812 (n = 8): Oral treatment with 120 mg / kg ABTL0812 (in 200 μl distilled water + 5% glycerol) 5 times a week; ip administration of 60 mg / kg gemcitabine (in 100 μl sterile water) twice a week; and ip administration of 5 mg / kg Nab-paclitaxel (in 230 μl 0.9% NaCl solution) twice a week. Injections were administered in separate areas of the abdomen as in the gemcitabine + Nab-paclitaxel group. ABTL0812 was always administered before chemotherapy in order to reduce the stress caused by ip injection.

[0558] The efficacy of treatment was assessed by measuring tumor volume three days per week. In addition, total body weight was monitored three days per week to test for treatment-related toxicity, in addition to visual inspection for signs of clinical pathological symptoms. At the end of the study, mice were euthanized by carbon dioxide inhalation, and death was further confirmed by cervical dislocation.

[0559] Results: We chose a suboptimal dose of chemotherapy (based on the bibliography) with the goal of not having a strong antitumor response that could be enhanced by ABTL0812, thereby reducing the chemotherapy dose and thus its undesirable adverse events. Fig.14 Shown is the progression of tumor volume in MiaPaca2-derived xenografts treated with different chemotherapy regimens (ABTL0812 or a combination of both). Fig.14A shows the tumor volume progression of xenografts treated with gemcitabine+Nab-Pac (compared with gemcitabine+Nab-Pac+ABTL0812 treatment) in addition to ABTL0812 and vehicle treatment. Compared with the vehicle group, only the triple combination Gem+Nab-Pac+ABTL0812 showed significant tumor volume reduction from the 22nd day, and maintained this statistical significance until the end of the study, with the highest tumor volume difference shown in the last five days of treatment, and in contrast to the Gem+Nab-Pac group that did not show a significant difference in tumor volume compared to the vehicle group. When comparing Gem+Nab-Pac+ABTL0812 with Gem+Nab-Pac, the triple combination significantly reduced tumor volume on the last day of treatment. It can also be observed that ABTL0812 administered alone showed a better response in tumor volume evolution during the first 20 days of treatment compared to the Gem+Nab-Pac group, which was similar during the rest of the study, but no significant differences were observed at any time point.

[0560] Fig.14 B shows the tumor volume progression of xenografts treated with gemcitabine (compared with gemcitabine+ABTL0812 treatment) in addition to ABTL0812 and vehicle treatment. Compared with the vehicle group, only the double combination Gem+ABTL0812 showed a significant tumor volume reduction from the 17th day, and maintained this statistical significance until the end of the study, in contrast to the gemcitabine group that did not show a significant difference in tumor volume compared to the vehicle group. When comparing Gem and Gem+ABTL0812, the double combination significantly reduced tumor volume on the last day of treatment. It can also be observed that compared with the Gem group, during the entire study period, ABTL0812 administered alone showed a better response in tumor volume evolution, but no significant difference was observed at any time point.

[0561] Total body weight of mice was monitored three times per week throughout the study. Fig.14 C shows the total body weight evolution of Gem+Nab-Pac and Gem+Nab-Pa+ABTL0812 in addition to the vehicle and ABTL0812 groups, and Fig.14 D shows the total body weight evolution of Gem and Gem+ABTL0812 in addition to the vehicle and ABTL0812 groups. No signs of toxicity were observed in any group in terms of body weight loss, and all groups showed weight gain throughout the study, indicating a lack of treatment-related toxicity. No additional clinical pathological signs were observed in any group. For further details, see Fig.14 .

[0562] Conclusion: This study was designed to determine the efficacy of ABTL0812 alone or in combination with SOC chemotherapy in the treatment of advanced pancreatic cancer. The combination of gemcitabine and Nab-paclitaxel or gemcitabine alone is the first choice treatment for most patients with advanced pancreatic cancer, therefore, we evaluated the enhancement of both treatments by combining with ABTL0812 and using suboptimal doses of chemotherapy (thereby reducing undesirable secondary effects). ABTL0812 enhanced both chemotherapies while reducing toxicity, showing the highest tumor volume reduction compared to vehicle and compared to chemotherapy alone. In addition, ABTL0812 administered alone showed efficacy similar to chemotherapy, and no clinical pathology or toxicity-related signs in terms of total body weight were observed in any treatment group. 2.9：

[0564] Efficacy study of ABTL0812 alone or in combination with cisplatin in a neuroblastoma xenograft model (cisplatin-sensitive)

[0565] Research reference: ABTL0812 notebook, pg 20-36

[0566] Study location: Vail d'Hebron Research Institute Pediatric Cancer Translational Research Laboratory

[0567] GLP Compliance: No

[0568] Test compound: ABTL0812 (Batch 002 / 2013)

[0569] Reference compound: Cisplatin (Sigma #C2210000)

[0570] Test system: Nu / nu female mice, SH-SY5Y cell line

[0571] Objective: To determine the efficacy of ABTL0812 alone or in combination with cisplatin in the neuroblastoma cell line SH-SY5Y.

[0572] Methods: Immunocompromised athymic NMRI-Foxn1 nu / Foxn1 nu Nude mice were injected subcutaneously with SH-SY5Y cells. This cell line has been genetically modified to express luciferase, which allows in vivo studies not only of tumor size but also of metastasis formation. 3The mice were randomly distributed into different treatment groups when the mean volume of ABTL0812 was measured. ABTL0812 was orally administered at 120 mg / kg daily. At the same time, we used cisplatin, a drug included in the chemo-induction phase of the treatment of neuroblastoma. Cisplatin was administered at 2 mg / kg ip twice a week (Wang et al., 2010). In addition, we studied the effect of combining ABTL0812 with the indicated doses of cisplatin.

[0573] result: A. Tumor size. This experiment shows that ABTL0812 inhibits tumor progression with an efficacy similar to that of cisplatin. After ten days of treatment, animals in the control group were sacrificed due to ethical issues related to tumor size. At this time, half of the animals in the treatment group were sacrificed to measure tumor weight, hematological parameters, and metastasis formation (see below). Animals for sacrifice were selected in these groups based on the statistical distribution of tumor size. Tumor weight measurements of the sacrificed mice confirmed the observation that the efficacy of ABTL0812 was similar to that of the standard of care cisplatin. Analysis of tumor volumes in the remaining animals in the treatment groups (approximately 5 mice per group) showed that the combination of ABTL0812 and cisplatin achieved long-term stabilization of tumor growth. Weight monitoring showed that treatment with ABTL0812 transiently induced a mild weight loss (<10%). However, this effect was restored after a few days. For further details, see the article. Fig.15 .

[0574] B. Security. Hematological analysis to assess the safety of treatment showed that ABTL0812 had no effect on hematocrit, but cisplatin induced anemia and decreased white blood cell counts (see table below). Cisplatin-associated anemia is a common side effect observed in patients treated with this chemotherapeutic drug (Wood and Hrushesky 1995).

[0575] Table. Hematological analysis of animals in the SH-SY5Y xenograft model. Blood was collected from animals at sacrifice and blood composition was determined using an automated analyzer. *p<0.05 ANOVA followed by Bonferroni.

[0576]

[0577] C. Metastasis formation.In order to study the effect of ABTL0812 in metastasis formation, we used the SH-SY5Y cell line transduced with a luciferase reporter vector. As described above, mice carrying xenograft tumors derived from this cell line were treated with ABTL0812, cisplatin or a combination of two drugs. After treatment for ten days, the animals were killed and the cells expressing luciferase in the lungs and liver were evaluated in vitro for metastasis. These analyses show that ABTL0812, whether a single agent or in combination with cisplatin, inhibits metastasis formation in these organs. On the contrary, compared with vehicle-treated animals, cisplatin alone has no significant effect on inhibiting metastasis formation.

[0578] Conclusions: ABTL0812 as a single agent has comparable efficacy to cisplatin while having a better safety profile in terms of hematological parameters. Interestingly, the combination of ABTL0812 with cisplatin stabilized tumor progression for a longer period of time. Additionally, ABTL0812 suppressed spontaneous metastasis formation in a neuroblastoma mouse model, whereas cisplatin did not. These data further support that ABTL0812 may have an enhanced therapeutic effect compared to current platinum-based chemotherapies. 2.10：

[0580] Efficacy of ABTL0812 in combination with doxorubicin in human triple-negative breast cancer (MDA-MB-231) xenografts in mice

[0581] Research reference: To be determined

[0582] Study location: UAB Competence Laboratory.

[0583] GLP Compliance: No

[0584] Test compound: ABTL0812 (Batch 002 / 2012)

[0585] Reference compound: Doxorubicin (sigma#D1515)

[0586] Test system: Nu / nu male mice

[0587] Objective: To investigate the antitumor activity of ABTL0812 alone and in combination with doxorubicin in human triple-negative breast cancer xenografts.

[0588] Methods: MDA-DB-231 cell lines were routinely cultured in DMEM 10% FBS, and cells in the exponential growth phase were harvested and counted for tumor inoculation. Mice were injected with 2.5x10 6MDA-DB-231 cells were suspended in 50 μl of growth medium without FBS and 50 μl of Matrigel (Corning #354234). Tumor volume was monitored 3 days per week and when tumors reached 100 mm 3 (between 50 and 150mm 3 Animals were homogenously distributed into four treatment groups that showed similar mean intra-group tumor volumes (excluding tumors less than 50 mm 3 Or more than 150mm 3 of tumors to minimize differences).

[0589] The treatment groups were:

[0590] - Vehicle group (n=7): Oral treatment with 200 μl water + 5% glycerol four days a week; and two ip injections of 100 μl saline solution once a week

[0591] -ABTL0812 (n=9): Oral administration of 200 μl 120 mg / kg ABTL0812 (resuspended in water + 5% glycerol) 5 times a week

[0592] - Doxorubicin (n=9): 100 μl 2 mg / kg doxorubicin treatment ip once a week

[0593] -ABTL0812+doxorubicin (n=9): 200 μl 120 mg / kg ABTL0812 orally, four days a week; and 100 μl 2 mg / kg doxorubicin ip, once a week

[0594] Results: The combination of ABTL0812 administered with doxorubicin showed the best in vivo antitumor effect in xenografts derived from MDA-DB-231 cells. ABTL0812 administered alone showed similar tumor volume reduction as doxorubicin alone, but the double combination of ABTL0812 and doxorubicin showed the highest tumor volume reduction, and the difference was significant at the end of the study. All treatment groups showed similar overall body weight evolution, indicating a lack of treatment-related toxicity. For further details, see the article Fig.16 .

[0595] Conclusion: As previously reported, ABTL0812 reduced tumor growth in a breast cancer xenograft model derived from MDA-DB-231 cells. In this model, ABTL0812 enhanced the antitumor activity of doxorubicin. Doxorubicin treatment is a common treatment option for breast cancer patients, so these results suggest that the combination of ABTL0812 plus doxorubicin may have clinical significance for the treatment of breast cancer patients. 2.11：

[0597] Efficacy of ABTL0812 in combination with gemcitabine and cisplatin in human cholangiocarcinoma (EGI-1) xenografts in mice

[0598] Research reference: ABT-EI

[0599] Study Location: Liver Disease Group, Biodonostia Health Research Institute

[0600] GLP Compliance: No

[0601] Test compound: ABTL0812 (Batch 002 / 2012)

[0602] Reference compounds: Gemcitabine hydrochloride (Sigma Aldrich G6423) and cisplatin (Sigma #C2210000)

[0603] Test system: Nu / nu male mice

[0604] Objective: To investigate the antitumor activity of ABTL0812 alone and in combination with gemcitabine and cisplatin in human cholangiocarcinoma xenografts. Gemcitabine and cisplatin combination is one of the reference treatments for cholangiocarcinoma.

[0605] Methods: EGI-1 cell line was routinely cultured in DMEM 10% FBS, and cells in exponential growth phase were harvested and counted for tumor inoculation. Mice were injected with 1x10 6 H1957 cells suspended in 50 μl of growth medium without FBS and 50 μl of Matrigel (Corning #354234). Tumor volume was monitored 3 days per week and when tumors reached 100 mm 3 (between 50 and 150mm 3 Animals were homogenously distributed into four treatment groups that showed similar mean intra-group tumor volumes (excluding tumors less than 50 mm 3 Or more than 150mm 3 of tumors to minimize differences).

[0606] The treatment groups were:

[0607] - Vehicle group (n=8): Oral treatment with 200 μl water + 5% glycerol four days a week; and two ip injections of 100 μl saline solution once a week

[0608] -ABTL0812 (n=8): Oral administration of 200 μl 120 mg / kg ABTL0812 (resuspended in water + 5% glycerol) 5 times a week

[0609] Gemcitabine / cisplatin (n=8): 100 μl 50 mg / kg gemcitabine treatment ip; and 100 μl 2 mg / kg cisplatin ip once a week

[0610] ABTL0812 + gemcitabine / cisplatin (n=8): 200 μl 120 mg / kg ABTL0812 treatment orally four days a week; and 100 μl 50 mg / kg gemcitabine ip and 100 μl 2 mg / kg cisplatin ip once a week

[0611] Results: The combination of ABTL0812 administered with gemcitabine and cisplatin showed the best in vivo antitumor effect in xenografts derived from EGI-1 cells. Administration of gemcitabine and cisplatin reduced tumor volume compared to the vehicle group, but without any statistically significant difference, in contrast to ABTL0812+gemcitabine and cisplatin treatment which showed a statistically significant tumor volume reduction compared to vehicle, ABTL0812 administered alone did not show a tumor volume reduction compared to vehicle until the last day of treatment, where it showed similar tumor volume to the chemotherapy group. For further details, see Fig.17 .

[0612] Conclusion: As previously reported, ABTL0812 reduced tumor growth in a cholangiocarcinoma xenograft model derived from EGI-1 cells. In this model, ABTL0812 enhanced the antitumor activity of gemcitabine and cisplatin administration. Gemcitabine and cisplatin treatment is a common treatment option for patients with cholangiocarcinoma, so these results suggest that the combination therapy of ABTL0812 plus gemcitabine and cisplatin may be clinically meaningful for the treatment of patients with cholangiocarcinoma.

[0613] Example 3 : Toxicity of combination with chemotherapy

[0614] Information on the toxicity of the combination of ABTL0812 with chemotherapeutic agents was obtained during efficacy studies in immunosuppressed mice. Specific toxicology studies have been performed on the combination of ABTL0812 + / - paclitaxel + / - carboplatin.

[0615] Research reference: N-02220

[0616] Study location: Vivotecnia (Madrid, Spain)

[0617] GLP study: No

[0618] Test compounds: ABTL0812 (Batch 001R / 2014), Paclitaxel (Aurovitas, Batch 68J5041), Carboplatin (Sigma-Aldrich, Batch LSBL7058v)

[0619] Test system: CD-1 female mice, 12 weeks old.

[0620] Objective: To determine the toxicological profile of ABTL0812 in combination with carboplatin and paclitaxel after two weeks of administration.

[0621] Methods: Forty-five female CD-1 mice were allocated by body weight stratification to one experimental group (AI) (5 animals per group), each group differing in the number of days of treatment or reference administration (paclitaxel and carboplatin).

[0622] The following table summarizes the various treatment groups. Note that the same administration schedule and doses as the previous efficacy study were used.

[0623] Table. Treatment groups used to evaluate the toxicity of ABTL0812, paclitaxel, carboplatin, and their combination in mice.

[0624]

[0625]

[0626] Safety assessments rely on observed mortality, local and systemic clinical signs, body weight, and food consumption recorded throughout the study. In addition, clinical pathology determinations (biochemistry and hematology) were performed before all animals were sacrificed. At the end of the observation period (one day after the last administration), all surviving animals were sacrificed and subjected to macroscopic autopsy. In addition, safety assessments were also based on the weight of selected target organs collected at the time of sacrifice.

[0627] Results: Repeated oral treatment with the test item and intraperitoneal treatment with the reference item resulted in no mortality. No local or systemic clinical signs related to the treatment were recorded.

[0628] During the study, a slight difference in average weight gain was observed in the animals. Except for the animals in Group A (control group) and Group C (treated with a certain dose of paclitaxel, once a week), most animals showed a trend of weight loss in the first four days of the study. However, during the entire study, no statistically significant difference in absolute weight gain was observed in most animals. Compared with several animal groups (Group A treated with vehicle, Group C treated with paclitaxel, Group E treated with a combination of paclitaxel and carboplatin), during the entire study, only the animals in Group F (orally administered ABTL0812, once a day, for two 5-day stages; and intraperitoneally administered paclitaxel and carboplatin, once a week (the first oral administration day of each stage)) had a significant reduction in weight gain.

[0629] Although statistical analysis could not be performed due to the small sample size, the estimated food consumption of animals in group A appeared to be higher than that of the remaining animals. Regarding clinical biochemical parameters, creatinine and triglyceride levels were lower in most groups compared to the control group. On the other hand, a statistically significant decrease in hematocrit levels (animals in group F) and a statistically significant decrease in platelet levels (animals in groups F and G) were observed in the groups treated with a combination of oral ABTL0812 administration and intraperitoneal paclitaxel and carboplatin compared to the control group. Although these values ​​were within the normal range, a therapeutic effect on clinical pathological parameters cannot be excluded.

[0630] Post-mortem macroscopic observation of all animals euthanized at the end of treatment did not reveal any changes that were considered relevant to the test item. White areas were observed on the liver of animal ID 32 and pigmentation was observed on the pancreas of animal ID 33. Otherwise, absolute and relative organ weights were similar between treatment groups.

[0631] The table below summarizes the most important findings from a safety perspective.

[0632] Table. Most relevant biochemical and hematological findings from toxicology studies of ABTL0812 and its combination with paclitaxel and carboplatin

[0633]

[0634] Conclusions: Taking into account the results obtained, it can be concluded that under the experimental conditions determined:

[0635] - Repeated administration of the oral test item ABTL0812 alone and in combination with weekly intraperitoneal administration of paclitaxel and carboplatin resulted in no mortality and was well tolerated as no local or systemic clinical signs indicative of toxicity were observed in any animal during the study.

[0636] - Most animals maintained their body weight throughout the study period. Statistically significant decreases were observed only in animals treated with the combination of the test item and the two reference items (also on the first day of each period).

[0637] - The food consumption of animals in group A appeared to be higher than that of almost all animals during the study period.

[0638] - Clinical pathology (biochemical and hematological parameters) in the treated groups showed statistically significant differences compared to the control group. Although most values ​​were within the normal range for animals of this strain and sex, a therapeutic effect cannot be excluded. The effect on platelet counts was due to carboplatin, but an additional slight decrease in platelet counts when the three experimental drugs were combined could not be ignored.

[0639] - No treatment-related effects were observed on gross necropsy findings and absolute / relative organ weights.

[0640] Example 4 : Combinations of ABTL0812 with different chemotherapeutic agents - human data

[0641] 4.1： A Phase I / II open-label study to evaluate the efficacy and safety of ABTL0812 in combination with paclitaxel and carboplatin in patients with advanced endometrial carcinoma or squamous NSCLC.

[0642] A Phase I / II clinical trial is being conducted in patients with advanced endometrial cancer or squamous non-small cell lung cancer. This is a multicenter open-label trial in which ABTL0812 is used orally at a starting dose of 1300 mg three times a day in combination with chemotherapy.

[0643] A. Purpose of the Test

[0644] Phase I primary endpoint: To evaluate the safety and tolerability of ABTL0812 plus paclitaxel + carboplatin as first-line treatment in patients with advanced or metastatic endometrial cancer or squamous NSCLC

[0645] Phase II primary endpoint: Evaluate the efficacy of ABTL0812 plus paclitaxel + carboplatin as first-line treatment in patients with advanced or metastatic endometrial cancer or squamous NSCLC

[0646] B. Research Design

[0647] This study was not randomized, and all enrolled patients received ABTL0812 in addition to paclitaxel + carboplatin. This period was divided into 2 phases:

[0648] Phase 1: ABTL0812 is administered in combination with chemotherapy.

[0649] Phase 2: After the SOC cycle, ABTL0812 as monotherapy from the start of Phase 1 for up to 12 months.

[0650] in conclusion:

[0651] These results suggest that the synergistic effects associated with the use of the ABTL0812 compound in combination with paclitaxel and carboplatin in patients with advanced endometrial cancer or squamous cell carcinoma also exist in humans, and in this sense the obtained human clinical trials are positive.

[0652] References

[0653] 1: EP2409963B1 (Lipopharma – filed in 2010)

[0654] 2: Erazo et al.; Clinical Cancer Research; 22(10), May 15, 2016

[0655] 3: News dated November 22, 2016 - published on the website of the applicant (AbilityPharma) 4: News dated December 14, 2016 - published on the website of the applicant (AbilityPharma)

Claims

1. A drug combination comprising: (A): a compound of the polyunsaturated fatty acid of the formula COOH-CHOH-(CH2)6-(CH=CH-CH2)2-(CH2)3-CH3 (ABTL0812), a pharmaceutically acceptable salt thereof or a combination thereof and (B): a chemotherapeutic compound, wherein compound (B) is at least one chemotherapeutic compound selected from the group consisting of: Cyclophosphamide; Melphalan; Docetaxel; Paclitaxel; Nab-paclitaxel; Carboplatin; Cisplatin; Oxaliplatin; Methotrexate Pemetrexed; Azathioprine; Capecitabine; Fluorouracil; Mercaptopurine; Gemcitabine; Bleomycin; Actinomycin; Vincristine; Vinblastine; Vinorelbine; Retinoic acid; Temozolomide; Daunorubicin Doxorubicin; Irinotecan; and Topotecan They are used for simultaneous, separate or sequential use in the treatment of cancer in human patients.

2. The pharmaceutical combination according to any one of the preceding claims, wherein compound (B) is at least one chemotherapeutic compound selected from the group consisting of: Docetaxel; Paclitaxel; Nab-paclitaxel; Carboplatin; Cisplatin; Oxaliplatin; Methotrexate Pemetrexed; Gemcitabine; Bleomycin Retinoic acid; Temozolomide; Doxorubicin; irinotecan; and Topotecan.

3. A pharmaceutical combination according to any one of the preceding claims, wherein compound (A) is the sodium salt of COOH-CHOH-(CH2)6-(CH=CH-CH2)2-(CH2)3-CH3 (ABTL0812).

4. The pharmaceutical combination according to any one of the preceding claims, wherein the cancer is at least one cancer selected from the group consisting of: Lung cancer; Non-small cell lung cancer; Small cell lung cancer; Squamous cell carcinoma; Adenocarcinoma; Endometrial cancer; Pancreatic cancer; Glioblastoma; Breast cancer; Head and neck cancer; neuroblastoma; and Biliary duct cancer.

5. The pharmaceutical combination according to any one of the preceding claims, wherein the cancer is at least one cancer selected from the group consisting of: Non-small cell lung cancer; Squamous cell carcinoma; Endometrial cancer; Pancreatic cancer; Glioblastoma; Breast cancer; neuroblastoma; and Biliary duct cancer.

6. A pharmaceutical combination according to any one of the preceding claims, wherein Compound (B) is docetaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma; Compound (B) is paclitaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma; Compound (B) is gemcitabine, and the cancer is pancreatic cancer; Compound (B) is carboplatin, and the cancer is endometrial cancer; Compound (B) is retinoic acid, and the cancer is neuroblastoma; Compound (B) is paclitaxel, and the cancer is breast cancer, preferably triple-negative breast cancer; Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma, preferably non-small cell lung squamous cell carcinoma; Compound (B) is paclitaxel and carboplatin, and the cancer is non-small cell lung adenocarcinoma; Compound (B) is pemetrexed and cisplatin, and the cancer is non-small cell lung adenocarcinoma; Compound (B) is paclitaxel, and the cancer is endometrial cancer; Compound (B) is paclitaxel and carboplatin, and the cancer is endometrial cancer; Compound (B) is paclitaxel and gemcitabine, and the cancer is pancreatic cancer; Compound (B) is gemcitabine, and the cancer is pancreatic cancer; Compound (B) is gemcitabine and Nab-paclitaxel, and the cancer is pancreatic cancer; Compound (B) is cisplatin and gemcitabine, and the cancer is neuroblastoma; Compound (B) is doxorubicin acid, and the cancer is breast cancer, preferably triple-negative breast cancer; Compound (B) is cisplatin and gemcitabine, and the cancer is cholangiocarcinoma; Compound (B) is gemcitabine, and the cancer is cholangiocarcinoma; Compound (B) is topotecan, and the cancer is neuroblastoma; Compound (B) is irinotecan, and the cancer is neuroblastoma; or Compound (B) is temozolomide and the cancer is neuroblastoma.

7. A pharmaceutical combination according to any one of the preceding claims, wherein Compound (B) is docetaxel, and the cancer is lung cancer, preferably non-small cell lung adenocarcinoma; Compound (B) is gemcitabine, and the cancer is pancreatic cancer; Compound (B) is carboplatin, and the cancer is endometrial cancer; Compound (B) is retinoic acid, and the cancer is neuroblastoma; Compound (B) is paclitaxel, and the cancer is breast cancer, preferably triple-negative breast cancer; Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma, preferably non-small cell lung squamous cell carcinoma; Compound (B) is paclitaxel and carboplatin, and the cancer is non-small cell lung adenocarcinoma; Compound (B) is pemetrexed and cisplatin, and the cancer is non-small cell lung adenocarcinoma; Compound (B) is paclitaxel and carboplatin, and the cancer is endometrial cancer; Compound (B) is paclitaxel and gemcitabine, and the cancer is pancreatic cancer; Compound (B) is gemcitabine, and the cancer is pancreatic cancer; Compound (B) is gemcitabine and Nab-paclitaxel, and the cancer is pancreatic cancer; Compound (B) is doxorubicin acid, and the cancer is breast cancer, preferably triple-negative breast cancer; Compound (B) is cisplatin and gemcitabine, and the cancer is cholangiocarcinoma; or Compound (B) is gemcitabine and the cancer is cholangiocarcinoma.

8. A pharmaceutical combination according to any one of the preceding claims, wherein Compound (B) is paclitaxel and carboplatin, and the cancer is advanced endometrial cancer, preferably advanced endometrial cancer; or Compound (B) is paclitaxel and carboplatin, and the cancer is squamous cell carcinoma.

9. The pharmaceutical combination according to claim 8, wherein compound (A) is administered orally.

10. The pharmaceutical combination according to claim 9, wherein the administered dose of compound (A) is a dose of 1200 mg to 1400 mg.

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