KR20190074946A - Therapeutic compositions for treating pancreatic cancer - Google Patents

Abstract

The present invention relates to a therapeutic composition for treating a pancreatic cancer, which comprises a cyclohexenone compound and at least one anticancer agent or a pharmaceutically acceptable salt, metabolite, a solvate or prodrug thereof.

Description

[0001] THERAPEUTIC COMPOSITIONS FOR TREATING PANCREATIC CANCER [0002]

The present invention relates to a therapeutic composition for treating pancreatic cancer.

Pancreatic cancer occurs when the pancreatic cells proliferate uncontrollably and form lumps. These cancer cells have the ability to invade other parts of the body. Pancreatic cancer is the fourth most frequent cause of cancer deaths in the West and is also the tenth most frequent cause of cancer deaths in Taiwan. Approximately 60% of pancreatic cancer occurs in the head of the body, and approximately 21% of the pancreas is invaded by the entire pancreas.

Pancreatic cancer can be divided into two general groups. In the vast majority (about 99%), they occur in the pancreas where they produce digestive enzymes known as exocrine components. There are several subtypes of exocrine pancreatic cancer, but their diagnosis and treatment are much more common. These few cancers occur in the hormone-producing (endocrine) tissues of the pancreas.

Surgery for treatment is only possible in about 1/5 (20%) of the new cases. Although CT scans are helpful, it may be difficult to determine whether or not the tumor can be completely removed ("resectability"), and tumors can not be successfully removed without damaging other essential tissues. It can only be made clear during. Even if surgery seems to be successful, cancer cells are often found at the margins of the removed tissue (the "periphery"). After the surgery, after a recovery period of one to two months, adjuvant chemotherapy using gemcitabine or 5-FU may be provided if the patient is well-suited. In patients who are not suitable for surgery for treatment, chemotherapy may be used to prolong life or improve quality of life.

의 구조를 갖는 화합물 및 하나 이상의 항암제, 또는 이들의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을 포함하는, 개체에서 췌장암을 치료하기 위한 조성물이며, 상기 식에서, X 및 Y는 각각 독립적으로 산소 또는 황이고; R은 수소 또는 C(=O)C₁-C₈ 알킬이며; R₁, R₂ 및 R₃은 각각 독립적으로 수소, 메틸 또는 (CH₂)_m-CH₃이고; R₄는 H, C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, 또는 C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, C₃-C₈ 사이클로알킬 및 C₁-C₈ 할로알킬로부터 선택되는 하나 이상의 치환기로 임의적으로 치환되는 아릴이며; n은 1 내지 12이다.In one embodiment, provided herein are compounds of formula

A composition for treating pancreatic cancer in a subject, comprising a compound having the structure of Formula I and at least one anticancer agent, or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof, wherein X and Y are each Is independently oxygen or sulfur; R is hydrogen or _{C (= O) C 1 -C} 8 alkyl; R ₁ , R ₂ and R ₃ are each independently hydrogen, methyl or (CH ₂ ) _m -CH ₃ ; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alky carbonyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C ₈ with one or more substituents selected from haloalkyl, optionally substituted aryl; n is from 1 to 12;

의 구조를 갖는 화합물, 및 하나 이상의 항암제, 또는 이들의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을 투여함을 포함하는, 개체에서 췌장암을 치료하는 방법이며, 상기 식에서, X 및 Y는 각각 독립적으로 산소 또는 황이고; R은 수소 또는 C(=O)C₁-C₈ 알킬이며; R₁, R₂ 및 R₃은 각각 독립적으로 수소, 메틸 또는 (CH₂)_m-CH₃이고; R₄는 H, C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, 또는 C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, C₃-C₈ 사이클로알킬 및 C₁-C₈ 할로알킬로부터 선택되는 하나 이상의 치환기로 임의적으로 치환되는 아릴이며; n은 1 내지 12이다.In another aspect, provided herein are methods of treating a subject in need of pancreatic cancer treatment,

A method for treating pancreatic cancer in a subject, comprising administering to the individual a compound having the structure of Formula I, and one or more anticancer agents, or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof, wherein X and Y is each independently oxygen or sulfur; R is hydrogen or _{C (= O) C 1 -C} 8 alkyl; R ₁ , R ₂ and R ₃ are each independently hydrogen, methyl or (CH ₂ ) _m -CH ₃ ; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alky carbonyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C ₈ with one or more substituents selected from haloalkyl, optionally substituted aryl; n is from 1 to 12;

의 구조를 갖는 화합물, 또는 그의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을 투여함을 포함하는, 겜시타빈, 파클리탁셀 또는 이들의 조합에 내성이거나 이들에 의해 치료되지 않거나 또는 이들에 대해 무응답인 췌장암을 앓는 환자를 치료하는 방법이며, 상기 식에서, X 및 Y는 각각 독립적으로 산소 또는 황이고; R은 수소 또는 C(=O)C₁-C₈ 알킬이며; R₁, R₂ 및 R₃은 각각 독립적으로 수소, 메틸 또는 (CH₂)_m-CH₃이고; R₄는 H, C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, 또는 C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, C₃-C₈ 사이클로알킬 및 C₁-C₈ 할로알킬로부터 선택되는 하나 이상의 치환기로 임의적으로 치환되는 아릴이며; n은 1 내지 12이다.In another aspect, provided herein is a method of treating a patient in need of pancreatic cancer,

Or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, which is resistant to, or is not resistant to, gemcitabine, paclitaxel, or a combination thereof. A method for treating a patient suffering from pancreatic cancer, wherein X and Y are each independently oxygen or sulfur; R is hydrogen or _{C (= O) C 1 -C} 8 alkyl; R ₁ , R ₂ and R ₃ are each independently hydrogen, methyl or (CH ₂ ) _m -CH ₃ ; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alky carbonyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C ₈ with one or more substituents selected from haloalkyl, optionally substituted aryl; n is from 1 to 12;

References by reference

All publications, patents, and patent applications referred to herein are hereby incorporated by reference to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated by reference.

The novel features of the invention are set forth with particularity in the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS The features and advantages of the invention will be better understood by reference to the following detailed description of the invention and the accompanying drawings, which illustrate exemplary embodiments in which the principles of the invention may be employed.
Figures 1a / 1b provide the results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with (1a) for 48 hours and (Ib) for 72 hours with Exemplified Compound 1. The cytotoxic activity was measured using MTT assay.
Figure 2a / 2b provides the results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with (2a) for 48 hours and (2b) for 72 hours with Erlotinib. The cytotoxic activity was measured using MTT assay.
Figure 3a / 3b provides the results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with 5-FU (3a) for 48 hours and (3b) for 72 hours. The cytotoxic activity was measured using MTT assay.
Figures 4a / 4b provide results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with gemcitabine (4a) for 48 hours and (4b) for 72 hours. The cytotoxic activity was measured using MTT assay.
Figures 5a / 5b provide the results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with irinotecan (5a) for 48 hours and (5b) for 72 hours. The cytotoxic activity was measured using MTT assay.
6a / 6b provides results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with oxaliplatin (6a) for 48 hours and (6b) for 72 hours. The cytotoxic activity was measured using MTT assay.
Figures 7a / 7b provide the results of cytotoxic activity in AsPC-1 pancreatic cancer cells treated with paclitaxel (7a) for 48 hours and (7b) for 72 hours. The cytotoxic activity was measured using MTT assay.
Figure 8 shows the effect of compound 1 with paclitaxel, gemcitabine, 5-FU, oxaliplatin, allotinib or irinotecan, the chemotherapeutic drugs for the cytotoxicity of AsPC-1 pancreatic cancer cell lines for 48 hours. The cytotoxic activity was measured using MTT assay. Values are mean SEM of survival rates. The different letters a through e represent a significant difference (P < 0.05) for various treatments.
Figure 9 shows the cytotoxic activity of compound 1 with each of the chemotherapeutic drugs paclitaxel, gemcitabine, 5-FU, oxaliplatin, allotinib and irinotecan. AsPC-1 pancreatic cancer cells were treated with anthraquinolone or a combination preparation for 72 hours. The cytotoxic activity was measured using MTT assay. Values are mean SEM of survival rates. Different letters (a to g) represent significant differences (P < 0.05) for various treatments.
Figures 10a-10c illustrate the dose effect of a combination of paclitaxel (Px), gemcitabine (Ge) and compound 1 (also known as anthroquinolone Aq) in pancreatic cancer cell lines. Plots show the combination of anthroquinolone and paclitaxel in AsPC-1 (Fig. 10a), CaPan-2 (Fig. 10b) and Panc-1 (Fig. 10c) pancreatic cancer cell lines, a combination of anthroquinol and gemcitabine, Lt; RTI ID = 0.0 &gt; Fa &lt; / RTI &gt; treated with a combination of Nanol and paclitaxel and gemcitabine. The x-axis represents the dose (μ mol / L) of the drug and the y-axis represents Fa, ie the fraction of affected (growth inhibiting) cells.
FIGS. 11A-C illustrate the results of analysis of the combination of paclitaxel, gemcitabine and compound 1 in pancreatic cancer cell lines for 72 hours. A combination of anthroquinolone (Aq) and paclitaxel (Px) in AsPC-1 (FIG. 11A), CaPan-2 (FIG. 11B) and Panc-1 (FIG. 11C) pancreatic cancer cell lines, a combination of anthroquinol and gemcitabine ), A CI plot for a combination of anthroquinolone and paclitaxel and gemcitabine. The x-axis represents Fa (fraction of affected cells) and the y-axis represents CI (combination index). CI = 1, < 1 and &gt; 1 represent additive, synergistic and antagonistic effects, respectively.
Figure 12 shows the results of mouse weight changes during chemotherapeutic drug and compound 1 in an AsPC-1 subcutaneous xenograft model during an anticancer activity test.
Figure 13 shows the effect of chemotherapeutic drugs on the AsPC-1 subcutaneous xenograft model and the anticancer activity of Compound 1. Tumor size was measured weekly and the mean relative tumor volume was plotted as a function of time after treatment initiation. Each point represents the mean tumor volume.
Figure 14 shows the effect of Compound 1 and a chemotherapeutic drug on the AsPC-1 subcutaneous xenograft model. The percentage of tumor volume was significantly reduced in the treatment group of the combination containing Compound 1 compared to the control (T1, corn oil).
FIG. 15 shows the results of analysis of tumor growth inhibition rate (TGI) of Compound 1 alone or chemotherapeutic drugs paclitaxel and gemcitabine and Compound 1 for growth of AsPC-1 tumor volume. Each column showed an average tumor growth inhibition rate. Values are mean ± SEM. The different subscripts represent a significant difference (P &lt; 0.05) between each treatment group.
Figure 16 shows a comparison of representative photographs of AsPC-1 pancreatic cancer xenografts before and after treatment with Compound 1 and / or chemotherapy drugs, respectively.
Figure 17 shows the results of comparing representative ultrasonic images of AsPC-1 pancreatic cancer xenografts before and after treatment with Compound 1 and / or chemotherapeutic drugs, respectively.
Figures 18a / 18b show images at the time of death (Figure 18a) and Figure (Figure 18b) of tumors resected from male nude mice with subcutaneous AsPC-1 pancreatic cancer xenograft after 28 days of treatment. Statistical significance was analyzed using ANOVA. ^* P <0.05 was considered statistically significant when compared with the control group (T1, corn oil group).
Figure 19 shows the results of the inhibitory effect of Compound 1 alone or chemotherapeutic drugs paclitaxel and gemcitabine and Compound 1 on the growth of AsPC-1 solid tumors. Each column showed an average tumor inhibition ratio (TIR). Values are mean ± SEM of TIR. The different subscripts represent a significant difference (P &lt; 0.05) between each treatment group.

Surgery for treatment is only possible in about 1/5 (20%) of the new cases. A high proportion of people (about 30 to 45%) still need to be treated for post-operative disease that has not been caused by cancer itself, although surgery for treatment no longer involves very high mortality rates that occurred until the 1980s . The most common complication of surgery is the difficulty of emptying the stomach. Supplemental chemotherapy using gemcitabine or 5-FU may be provided after surgery, if adequate for the patient, after a recovery period of one to two months. Gemcitabine was approved by the US Food and Drug Administration (FDA) in 1997 after reporting improvements in quality of life and a 5-week improvement in median survival in patients with clinically advanced pancreatic cancer. Chemotherapy with gemcitabine alone has been around for a decade, as multiple experiments with other drugs failed to demonstrate significantly better results.

FOLFIRINOX chemotherapy using four drugs has been found to be more effective than gemcitabine, but has significant side effects and is therefore suitable only for those who exhibit excellent performance. This is also the case for protein-bound paclitaxel (nab-paclitaxel) approved by the FDA in 2013 for use with gemcitabine in pancreatic cancer. However, changes over the last few years have only increased survival time by a few months.

A method of treating pancreatic cancer in an individual, including administering a cyclohexanone compound to a subject in need thereof, is disclosed in U.S. Patent No. 8,236,860. Stereotactic transplantation An exemplary compound, anthroquinol, was studied for the treatment of pancreatic cancer using the PANC-1 human pancreatic cancer xenograft model. Four groups of mice were treated with 30 mg / kg, 60 mg / kg, 90 mg / kg and vehicle, respectively. Treatment with 30, 60 and 90 mg / kg of anthroquinolone produced effective anticancer activity with statistically significant mean tumor volume and tumor weight at all three dose levels compared to vehicle control. Although antroquinolone has been found to be a good candidate for the treatment of pancreatic cancer, it has not been considered for use in any concomitant therapy.

의 구조를 갖는 화합물, 및 겜시타빈, 파클리탁셀 또는 이들의 조합 같은 하나 이상의 항암제, 또는 이들의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을 포함하는 병행 요법에 상승 효과가 존재하는 것으로 밝혀졌으며, 상기 식에서, X 및 Y는 각각 독립적으로 산소 또는 황이고; R은 수소 또는 C(=O)C₁-C₈ 알킬이며; R₁, R₂ 및 R₃은 각각 독립적으로 수소, 메틸 또는 (CH₂)_m-CH₃이고; R₄는 H, C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, 또는 C₁-C₈ 알킬, C₂-C₈ 알케닐, C₂-C₈ 알키닐, C₃-C₈ 사이클로알킬 및 C₁-C₈ 할로알킬로부터 선택되는 하나 이상의 치환기로 임의적으로 치환되는 아릴이며; n은 1 내지 12이다. 일부 실시양태에서, 이러한 병행 요법은 면역 치료제를 투여함을 추가로 포함한다.However, unexpectedly,

And a synergistic effect in a combination therapy comprising one or more anticancer agents such as gemcitabine, paclitaxel, or a combination thereof, or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof, Wherein X and Y are each independently oxygen or sulfur; R is hydrogen or _{C (= O) C 1 -C} 8 alkyl; R ₁ , R ₂ and R ₃ are each independently hydrogen, methyl or (CH ₂ ) _m -CH ₃ ; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alky carbonyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C ₈ with one or more substituents selected from haloalkyl, optionally substituted aryl; n is from 1 to 12; In some embodiments, the concurrent therapy further comprises administering an immunotherapeutic agent.

의 구조를 갖는 화합물, 및 하나 이상의 항암제(실시예 1 내지 3 참조)를 포함하는, 개체에서 췌장암을 치료하기 위한 조성물이다. 사이클로헥세논 화합물은 일부 실시양태에서는 천연 생성물의 추출물로부터 수득되며, 일부 다른 실시양태에서는 합성에 의해 제조된다. 예컨대 미국 특허 제 9365481 호 참조.In some embodiments, provided herein are compounds of formula

, And at least one anticancer agent (see Examples 1 to 3). Cyclohexenone compounds are obtained from extracts of natural products in some embodiments, and by synthesis in some other embodiments. See, for example, U.S. Patent No. 9365481.

In some embodiments, the at least one anticancer agent is selected from the group consisting of gemcitabine, paclitaxel, dirubicin / cytarabine, ethoside phosphate, glivec (imatinib), temozolomide, bortezomib, retrosol, cetuximab, , Nab-paclitaxel, docetaxel, allotinib, femetrexed, femetrexed / carboplatin, paxilixaxel / carboplatin, letrozole / cyclophosphamide, temsirolimus, bevacizumab / temsirolimus PF-05212384 / irinotecan, AZD2171, PF-04691502, cyclophosphamide, cisplatin, cytarabine / daunorubicin, ticilolimus, alloty Nepheline / temsirolimus, capecitabine, tamoxifen, bortezomib, trastuzumab, docetaxel / capecitabine, trastuzumab / tipiparnib, tifiparnib / gemcitabine, tetecane or combinations thereof Including . In certain embodiments, the at least one anticancer agent is gemcitabine, paclitaxel, or a combination thereof. In certain embodiments, the at least one anticancer agent is a combination of gemcitabine and paclitaxel.

For example, paclitaxel is classified as "plant alkaloids", "taxanes" and "microtubule inhibitors". It is used to treat various types of cancer such as ovarian cancer, breast cancer, lung cancer, Kaposi sarcoma, cervical cancer and pancreatic cancer. Albumin-conjugated paclitaxel (also known as abraxane, nab-paclitaxel) is an alternative formulation in which paclitaxel is bound to albumin nano-particles. Paclitaxel has several common properties, including nausea and vomiting, loss of appetite, change in taste, tapering or brittle hair, pain in the arm or leg joint lasting from two to three months, nail color change, It is known to have side effects. Paclitaxel is one of several cytoskeletal drugs targeting tubulin. Paclitaxel-treated cells have defects in spindle assembly, chromosome segregation and cell division. Unlike other tubulin-targeting drugs, such as colchicine, which inhibit microtubule assembly, paclitaxel stabilizes the microtubule polymer and protects it from degradation.

Another exemplary anticancer drug, gemcitabine, is a nucleoside analog used as a chemotherapeutic for the treatment of various types of cancer, including breast, ovarian, non-small cell lung, pancreatic and bladder cancers. Common side effects include myelosuppression, liver and kidney problems, nausea, fever, rash, shortness of breath, and hair loss. Use during pregnancy can harm the fetus. Gemcitabine belongs to the class of nucleoside analogs of drugs. It works by blocking the formation of new DNA and killing cells.

In some embodiments, there is provided a method of treating pancreatic cancer in a subject, comprising administering to a subject in need thereof a concurrent therapy composition as described herein. In some embodiments, a concurrent therapy composition as described herein for use in treating pancreatic cancer in an individual is provided in a therapeutically effective amount.

의 구조를 갖는 화합물, 또는 그의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을, 췌장암 치료가 필요한 환자에게 투여함을 포함하는, 겜시타빈, 파클리탁셀 또는 이들의 조합에 대해 내성이거나 이들에 의해 치료되지 않거나 또는 이들에 대해 무응답인 췌장암 환자를 치료하는 방법이 제공된다. 일부 실시양태에서, 방법은 또한 면역 치료제를 투여함을 추가로 포함한다. 일부 실시양태에서는, 겜시타빈, 파클리탁셀 또는 이들의 조합에 대해 내성이거나 이들에 의해 치료되지 않거나 또는 이들에 대해 무응답인 췌장암 환자의 치료에 사용하기 위한, 화학식

의 구조를 갖는 화합물, 또는 그의 약학적으로 허용가능한 염, 대사산물, 용매화물 또는 전구약물을 치료 효과량으로 제공한다. In some embodiments,

Or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, for the treatment of pancreatic cancer, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of formula There is provided a method of treating a pancreatic cancer patient who has not been treated or has not responded to them. In some embodiments, the method further comprises administering an immunotherapeutic agent. In some embodiments, for use in the treatment of patients with pancreatic cancer who are resistant to, or not responding to, gemcitabine, paclitaxel, or a combination thereof,

Or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, in a therapeutically effective amount.

In certain embodiments, the cancer is selected from the group consisting of gemcitabine, paclitaxel, dirubicin / cytarabine, ethoside phosphate, gleevec, temozolomide, bortezomib, letrozole, cetuximab, bevacizumab, nab-paclitaxel, docetaxel, Pemetrex / Carboplatin, Letrozole / Cyclophosphamide, Temsirolimus, Bevacizumab / Temsilolorimus, Filimovab, RAD001, Pemetrexed / Carboplatin, Caffeine, cisplatin, cytarabine / daunorubicin, tallirolimus, allotinib / temsirolimus, caffeine, caffeine, Pf-05212384 / Irinotecan, AZD2171, PF-04691502 Or resistant to a drug selected from the group consisting of tamoxifen, tamoxifen, bortezomib, trastuzumab, docetaxel / capecitabine, trastuzumab / tipiparnib, tipiparnib / gemcitabine,Or a non-responsive to, or for those not treated by. In certain embodiments, the cancer is resistant to, or unresponsive to, gemcitabine or paclitaxel. In certain embodiments, the cancer is resistant to, or unresponsive to, the combination of gemcitabine and paclitaxel.

의 구조를 갖는 사이클로헥세논 화합물은 임의의 적합한 출발 물질로부터 합성에 의해 또는 반-합성에 의해 제조된다. 다른 실시양태에서, 사이클로헥세논 화합물은 발효 등에 의해 제조된다. 예를 들어, 화합물 1, 3 및 4는 유기 용매 추출물로부터 단리되거나, 또는 합성에 의해 또는 반-합성에 의해 제조된다. 비제한적인 예시적인 화합물이 아래에 예시된다.In some embodiments,

Is prepared from any suitable starting material by synthesis or by semi-synthesis. In another embodiment, the cyclohexenone compound is prepared by fermentation or the like. For example, compounds 1, 3 and 4 are isolated from organic solvent extracts, or prepared synthetically or semi-synthetically. Non-limiting exemplary compounds are illustrated below.

In some embodiments, R is hydrogen, C (= O) C ₃ H ₇ , C (= O) C ₂ H _5, or C (= O) CH ₃ . In some embodiments, R &lt; _{1 &gt;} is hydrogen or methyl. In certain embodiments, R ₂ is hydrogen, methyl, ethyl, propyl, butyl, pentyl, or hexyl. In some embodiments, R ₃ is hydrogen, methyl, ethyl, propyl, butyl, pentyl or hexyl. In some embodiments, R &lt; ₄ &gt; is hydrogen. In some embodiments, R ₄ is C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl; Which is optionally substituted by one or more substituents selected from C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl and C ₁ -C ₈ haloalkyl, / RTI &gt; In certain embodiments, R ₄ is CH ₂ CH = C (CH ₃ ) ₂ . In certain embodiments, the compound is

이다.

to be.

In some embodiments, the method of treating pancreatic cancer described herein comprises administering an immunotherapeutic agent. An immunotherapeutic agent includes, but is not limited to, any live immune cell that can be administered to a patient, and / or an antibody specific for a target cell (e.g., a cancer cell, such as a pancreatic cancer cell). Preferably, the immunotherapeutic agent is an NK cell or T cell, or a modification or derivative thereof.

Immunotherapy

Immunotherapy is "treatment of a disease by inducing, enhancing or inhibiting an immune response ". Immunotherapy designed to attract or amplify the immune response is classified as active immunotherapy, while immunotherapy that reduces or suppresses it is classified as inhibitory immunotherapy.

In some embodiments, the immunotherapeutic agent is an anti-cancer antibody. Non-limiting examples include, but are not limited to, Trastuzumab (Herceptin®), Bevacizumab (Avastin®), Cetuximab [Erbitux®], Paneytumamab [Vectibix®] , Yupilimu Mab [Yervoy®], Rituximab [Rituxan®], Alememuzo Marb [Campath®], Opatomumab [Arzerra®], Gem Mylotarg ®, Bentuximabbototin [Adcetris ®], ⁹⁰ γ-iburtomomvitizuketane (Zevalin ®), ¹³¹ (Anti-PD-1) antibodies such as I-Tocitumomab [Bexxar®], nibolumab, pembrolium jumab, and the like.

Specific pharmacy terms and medical terms

Unless otherwise stated, the following terms used herein, including the specification and claims, have the definitions given below. It should be understood that the singular forms used herein and in the appended claims include plural referents unless the context clearly dictates otherwise. Unless otherwise indicated, conventional methods of mass spectrometry, NMR, HPLC, protein chemistry, biochemistry, recombinant DNA techniques and pharmacology are utilized. In this application, the use of "or" or "and" means "and / or" unless otherwise stated. In addition, the use of the terms "including, " and " includes," The compartment headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

An "alkyl" group refers to an aliphatic hydrocarbon group. The alkyl group may be a saturated alkyl group (meaning no carbon-carbon double bond or no carbon-carbon triple bond), or the alkyl group may be an unsaturated alkyl group (containing at least one carbon-carbon double bond or a carbon- Containing &quot;). The alkyl residues may be branched, whether saturated or unsaturated, or may be linear.

"Alkyl" groups may have from 1 to 12 carbon atoms. (Wherever it appears herein, a numerical range such as "1 to 12" refers to each integer within a given range, for example, "1 to 12 carbon atoms" Means that the alkyl group can be composed of up to 12 carbon atoms such as 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc. This definition is also applicable to the case of the term "alkyl" do). The acyl group of the compounds described herein may be referred to as "C ₁ -C ₁₂ alkyl", "C ₁ -C ₈ alkyl" or a similar designation. By way of example, "C ₁ -C ₈ alkyl" indicates that there are 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms in the alkyl chain. In one embodiment, the alkyl is selected from the group consisting of methyl, ethyl, propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, and tert-butyl. Typical alkyl groups include but are not limited to methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tertiary butyl, pentyl, neopentyl, hexyl, allyl, , Cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, and the like. In one embodiment, the alkyl is C ₁ -C ₈ alkyl.

The term "alkylene" refers to a divalent alkyl radical. Any of the aforementioned monovalent alkyl groups may be alkylene formed by removing a second hydrogen atom from the alkyl. In one embodiment, the alkylene is C ₁ -C ₁₂ alkylene. In another embodiment, the alkylene is C ₁ -C ₈ alkylene. Typical alkylene groups are _{-CH 2 -, -CH (CH 3} ) -, -C (CH 3) 2 -, -CH 2 CH 2 -, -CH 2 CH (CH 3) -, -CH 2 C (CH 3 ) ₂ -, -CH ₂ CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ CH ₂ -, and the like.

The term "aryl &quot;, as used herein, refers to an aromatic ring in which each of the atoms forming the ring is a carbon atom. Aryl rings are formed by 5, 6, 7, 8, 9 or more carbon atoms. The aryl group is optionally substituted. In one embodiment, the aryl is phenyl or naphthalenyl. In one embodiment, the aryl is phenyl. In one embodiment, the aryl is C ₆ -C ₁₀ aryl. Depending on the structure, the aryl group may be a single radical or a divalent radical (i.e., an arylene group). In one embodiment, the arylene is C ₆ -C ₁₀ arylene. Examples of arylene include, but are not limited to, phenyl-1,2-ene, phenyl-1,3-ene and phenyl-1,4-ene.

The term "aromatic" refers to a planar ring having a non-local pi-electron system containing 4n + 2 (where n is an integer) electrons. An aromatic ring can be formed from 5, 6, 7, 8, 9, 10 or more atoms. The aromatic compound is optionally substituted. The term "aromatic" includes both carbocyclic aryl ("aryl" such as phenyl) and heterocyclic aryl (or "heteroaryl" or "heteroaromatic") groups such as pyridine. The term includes monocyclic or fused ring cyclic (i. E., Rings that share adjacent pairs of carbon atoms) groups.

The term "halo" or alternatively "halogen" or "halide" means fluoro, chloro, bromo or iodo.

The term "alkenyl &quot;, as used herein, refers to straight, branched or cyclic (in this case" cycloalkenyl &quot;) radicals containing from two to ten carbons and containing at least one carbon- &Quot; or "hydrocarbons &quot;). The alkenyl groups of the compounds described herein may be referred to as "C ₂ -C ₁₀ alkenyl &quot;," C ₂ -C ₈ alkenyl " By way of example, "C ₂ -C ₈ alkenyl" indicates that there are 2, 3, 4, 5, 6, 7 or 8 carbon atoms in the alkenyl chain. In some embodiments, depending on the structure, the alkenyl group is a monovalent radical or a divalent radical (i.e., an alkenylene group). In some embodiments, the alkenyl group is optionally substituted. Illustrative examples of alkenyl are ethenyl, 2-propenyl, 2-methyl-2-propenyl, 3-butenyl, 4- pentenyl, 5- hexenyl, 2- Hexenyl, heptenyl, and 3-secenyl.

As used herein, the term "alkynyl" refers to straight, branched, or cyclic (in this case, "cycloalkenyl &quot;) containing from two to ten carbons and containing at least one carbon- (Also known as " Neal ") hydrocarbons. In some embodiments, depending on the structure, the alkynyl group is a monovalent radical or a divalent radical (i.e., an alkynylene group). The alkynyl group of the compounds described herein may be referred to as "C ₂ -C ₁₀ alkynyl", "C ₂ -C ₈ alkynyl" or a similar designation. As used herein, the term &quot; C ₂ -C ₈ alkynyl "means that there are 2, 3, 4, 5, 6, 7 or 8 carbon atoms in the alkynyl chain. In some embodiments the alkynyl group is optionally substituted. Exemplary examples of the Niet include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, and the like.

The term "cycloalkyl ", as used herein, refers to monocyclic or polycyclic radicals containing only carbon and hydrogen and includes saturated or partially unsaturated or fully unsaturated radicals. The cycloalkyl group includes a group having 3 to 10 ring atoms. Representative examples of cycloalkyl groups include, but are not limited to, the following moieties:

In some embodiments, depending on the structure, the cycloalkyl group is a monovalent radical or a divalent radical (e. G., A cycloalkylene group).

The term "haloalkyl", "haloalkenyl", "haloalkynyl" and "haloalkoxy" as used herein includes alkyl, alkenyl, alkynyl, and alkoxy structures in which one or more hydrogen is replaced by a halogen atom. In certain embodiments in which two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms are all identical to each other. In another embodiment where two or more hydrogen atoms are replaced by halogen atoms, the halogen atoms are not all identical to one another. The terms "fluoroalkyl" and "fluoroalkoxy" include haloalkyl and haloalkoxy groups, wherein each halo is fluoro. In certain embodiments, the haloalkyl is optionally substituted.

The term "acceptable" as used herein with reference to a formulation, composition or component means that it does not have a persistent adverse effect on the systemic health of the individual being treated.

Antrodia is a fungal genus of the family Meripilaceae. Antrodia species typically have a fruiting body that is flattened or spread on the growing surface (the fur- ther layer is exposed to the outside); The edges can be bent to form narrow parentheses. Most species are found in temperate and boreal forests and cause sclerotia.

The term "carrier" as used herein refers to a relatively non-toxic chemical compound or agent that facilitates incorporation of the compound into cells or tissues.

The term "co-administration ", as used herein, is intended to encompass administration of a selected therapeutic agent to a single patient and is intended to encompass a therapeutic regimen wherein the agent is administered by the same or different routes of administration, or simultaneously or at different times.

The term "diluent" refers to a chemical compound used to dilute a compound of interest prior to delivery. Diluents are also used to stabilize the compounds, since they can provide a more stable environment. Salts dissolved in a buffered solution, including but not limited to phosphate buffered saline solutions, which may also provide pH control or maintenance, are used in the art as diluents.

The term " effective amount "or" therapeutically effective amount ", as used herein, refers to a sufficient amount of the agent or compound to be administered that alleviates to some extent at least one of the symptoms of the disease or condition being treated. The result may be a reduction and / or alleviation of the symptoms, symptoms or causes of the disease, or any other desired change in the biological system. For example, an "effective amount" for therapeutic use is an amount of a composition comprising a compound disclosed herein, which is necessary to provide a clinically significant reduction in a disease symptom. An appropriate "effect" amount in any individual case may be determined using techniques such as dose escalation studies.

"Metabolite" of a compound disclosed herein is a derivative of a compound that is formed when a compound is metabolized. The term "active metabolite" refers to a biologically active derivative of a compound that is formed when a compound is metabolized. As used herein, the term "metabolized " refers to the sum of the processes by which a particular component is altered by an organism, including, but not limited to, reactions catalyzed by hydrolysis and enzymes. Thus, enzymes can create specific structural changes in compounds. For example, cytochrome P450 promotes a variety of oxidation and reduction reactions, while uridine diphosphate glucuronyltransferase catalyzes the oxidation and reduction of aromatic glucuronic acid molecules with aromatic alcohols, aliphatic alcohols, carboxylic acids, Promote the transmission of the crossroads. Metabolites of the compounds disclosed herein are optionally identified by administration of the compound to the host and analysis of tissue samples from the host or by in vitro incubation of the hepatocyte and the compound and analysis of the resulting compound.

The term "pharmaceutical combination" as used herein means a product formed by mixing or coalescing more than one active ingredient, and includes fixed combinations and non-fixed combinations of active ingredients. The term "fixed combination" refers to the simultaneous administration of the active ingredient, e.g., a compound (i. E., The cyclohexenone compound described herein) and co-agent, both singly or in a single dosage form, to a patient. The term "non-fixed combination" means that the active ingredient, such as the compound (i. E., The cyclohexenone compound described herein), and the co-agent are administered separately as separate entities simultaneously, either consecutively or without specific time interval restrictions And such administration provides an effective level of both compounds in the body of the patient. The latter also applies to cocktail therapy, for example the administration of three or more active ingredients.

The term "pharmaceutical composition" refers to a mixture of a compound (i. E., The cyclohexenone compounds described herein) with other chemical ingredients such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents and / or excipients. The pharmaceutical composition enables administration of the compound to an organism. A number of techniques exist in the art for administering the compounds, including, but not limited to, intravenous, oral, aerosol, parenteral, eye, pulmonary and topical administration.

The term "individual" or "patient" includes mammals. Examples of mammals include any member of the mammalian river, e. Non-human primates such as chimpanzees, and other apes and monkey species; Breeding animals such as cows, horses, sheep, goats and pigs; Rabbits, livestock such as dogs and cats; But are not limited to, laboratory animals including rodents such as rats, mice and guinea pigs. In one embodiment, the mammal is a human.

The term " treating, "" treating," or "treating ", as used herein, refers to alleviating, ameliorating or ameliorating one or more symptoms of a disease or disorder, preventing additional symptoms, To prevent the onset of the disease or illness, to alleviate the disease or illness, to cause the degeneration of the disease or illness, to alleviate the condition caused by the disease or illness, or to prevent the symptoms of the illness or disease and / .

Routes of administration and dosage

Suitable routes of administration include, but are not limited to, oral, intravenous, rectal, aerosol, parenteral, eye, pulmonary, transmucosal, transdermal, vaginal, ear, nose and topical administration. Also, by way of example, parenteral delivery includes intramuscular, subcutaneous, intravenous, intraspinal injection, and intraspinal instillation, direct intracerebral, intraperitoneal, intra-lymphatic, and intranasal injection.

In certain embodiments, the compounds described herein are administered locally rather than systemically, for example by direct injection of the compound into the organs, often with depot or slow release formulations. In a specific embodiment, the long acting formulations are administered by implantation (e. G., Subcutaneously or intramuscularly) or by intramuscular injection. Further, in another embodiment, the drug is delivered into a targeted drug delivery system, e.g., into a liposome coated with an organ-specific antibody. In such embodiments, the liposomes are targeted to the organs and selectively absorbed by the organs. In another embodiment, the compounds described herein are provided in the immediate release dosage form, in the sustained release dosage form, or in the form of an intermediate release dosage form. In another embodiment, the compounds described herein are administered locally.

In some embodiments, the cyclohexenone compound or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof is administered parenterally or intravenously. In another embodiment, the cyclohexenone compound, or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, is administered by injection. In another embodiment, the cyclohexenone compound, or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, is administered orally.

If the condition of the patient is not improved, the compound may be administered over a long period of time, including the entire period of the patient's life, to alleviate or otherwise inhibit or limit the symptoms of the patient's disease or disease, . If the condition of the patient is not improved, the compound may be administered continuously or at the discretion of the physician, or the compound administration is temporarily suspended (i.e., "drug resting") for a certain period of time.

The above-mentioned ranges are merely suggestions, since it is not uncommon to have a large number of variables associated with individual therapies and to deviate significantly from these recommended values. Such dosages will depend on a variety of variables including, but not limited to, the activity of the compound employed, the disease or condition being treated, the mode of administration, the requirements of the individual subject, the severity of the disease or disorder being treated, . &Lt; / RTI &gt;

또는

이다.Standard pharmaceuticals in cell culture media or laboratory animals including, but not limited to, determining the LD ₅₀ (the dose lethal to 50% of the population) and the ED ₅₀ (the dose therapeutically effective in 50% of the population) By the procedure, the toxicity and therapeutic efficacy of such therapeutic regimens can be determined. The dose ratio between toxic and therapeutic effects is the therapeutic index, which can be expressed as the ratio between LD ₅₀ and ED ₅₀ . Compounds that exhibit a high therapeutic index are preferred. The range of dosages for use in humans may be expressed using data obtained from cell culture assays and animal studies. The dosage of such compounds is preferably in the range of circulating concentrations that include the ED ₅₀ with minimal toxicity. The dosage can vary within this range depending on the dosage form employed and the route of administration utilized. In certain embodiments, the cyclohexenone compound is

or

to be.

In some embodiments, the compounds described herein are formulated with a pharmaceutical composition. In certain embodiments, the pharmaceutical compositions are formulated in a conventional manner using one or more physiologically acceptable carriers, including excipients and adjuvants, which facilitate the processing of the active compound into a pharmaceutical agent that can be used pharmaceutically. The appropriate formulation depends on the route of administration chosen. Any of the pharmaceutically acceptable techniques, carriers and excipients suitable for incorporation into the pharmaceutical compositions described herein are used: Remington : The Science and Practice of Pharmacy , 19th Edition (Easton, PA: Mack Publishing Company, 1995); Hoover, John E., Remington ' s Pharmaceutical Sciences , Mack Publishing Co., Easton, Pa. 1975; Liberman, HA and Lachman, L. Ed., Pharmaceutical Dosage Forms , Marcel Decker, New York, NY, 1980; And Pharmaceutical Dosage Forms and Drug Delivery Systems , Seventh Edition [Lippincott Williams & Wilkins (1999).

Provided herein are pharmaceutical compositions comprising a compound (i. E., A cyclohexenone compound as described herein) and a pharmaceutically acceptable diluent (s), excipient (s) or carrier (s). In certain embodiments, the disclosed compounds are administered as a pharmaceutical composition in which the compound (i. E., The cyclohexenone compound described herein) is combined with other active ingredients as a concurrent therapy. All combinations of the active ingredients described herein below and the concurrent therapy portion are included herein. In a specific embodiment, the pharmaceutical composition comprises one or more compounds (i. E., The cyclohexenone compounds described herein).

The pharmaceutical compositions used herein refer to a mixture of the compound (i. E., The cyclohexenone compounds described herein) with other chemical ingredients such as carriers, stabilizers, diluents, dispersants, suspending agents, thickening agents and / or excipients. In certain embodiments, the pharmaceutical composition enables administration of the compound to the organism. In some embodiments, in practicing the therapeutic method or use provided herein, a therapeutically effective amount of the compound (i. E., The cyclohexenone compound described herein) is administered to the mammal having the disease or condition to be treated with the pharmaceutical composition . In a specific embodiment, the mammal is a human. In certain embodiments, the therapeutically effective amount will depend on the severity of the disease, the age and relative health of the subject, the efficacy of the compound employed and other factors. The compounds described herein are used alone or in combination with one or more therapeutic agents.

In one embodiment, the compound (the cyclohexenone compound described herein) is formulated in an aqueous solution. In a specific embodiment, the aqueous solution is selected from physiologically compatible buffering agents such as, for example, Hank's solution, Ringer's solution or physiological saline buffer. In another embodiment, the compound (i. E., The cyclohexenone compound described herein) is formulated for transmucosal administration. In a specific embodiment, the light transmucosal formulation comprises a penetrant suitable for the barrier to be penetrated. In another embodiment where the compounds described herein are combined with other parenteral injections, suitable formulations include aqueous or non-aqueous solutions. In a specific embodiment, such solutions comprise physiologically compatible buffering agents and / or excipients.

In another embodiment, the compounds described herein are formulated for oral administration. Compounds described herein, including the compounds (i. E., The cyclohexenone compounds described herein), are formulated by incorporating the active compound with, for example, a pharmaceutically acceptable carrier or excipient. In various embodiments, the compounds described herein are formulated in an oral dosage form including, for example, tablets, powders, pills, dragees, capsules, liquids, gels, syrups, elixirs, slurries,

In certain embodiments, the pharmaceutical preparations for oral use are prepared by mixing one or more solid excipients with one or more of the compounds described herein, optionally milling the resulting mixture, adding suitable auxiliaries, if necessary, To obtain tablets or dragee cores. Suitable excipients specifically include fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; Cellulose preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth gum, methylcellulose, microcrystalline cellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose; Or other materials such as polyvinylpyrrolidone (PVP or povidone) or calcium phosphate. In a specific embodiment, the disintegrant is optionally added. Disintegrants include, for example, cross-linked croscarmellose sodium, polyvinylpyrrolidone, agar, or alginic acid, or salts thereof such as sodium alginate.

In one embodiment, dosage forms such as dragee cores and tablets are provided with one or more suitable coatings. In a specific embodiment, a concentrated solution is used to coat the dosage form. The sugar solution optionally contains additional ingredients such as, for example, gum arabic, talc, polyvinyl pyrrolidone, carbo-gel gel, polyethylene glycol and / or titanium dioxide, a lacquer solution, and a suitable organic solvent or solvent mixture. Dyes and / or pigments are also optionally added to the coating for identification purposes. In addition, dyes and / or pigments are optionally employed to characterize different combinations of active compound doses.

In certain embodiments, a therapeutically effective amount of one or more of the compounds described herein is combined with another oral dosage form. Oral dosage forms include push-fit capsules made of gelatin as well as soft, sealed capsules made of gelatin and plasticizers (e.g., glycerol or sorbitol). In a specific embodiment, the press-fit capsules contain the active ingredient mixed with one or more fillers. The filler includes, for example, a lubricant such as lactose, starch, and / or a lubricant such as talc or magnesium stearate, and optionally a stabilizer. In another embodiment, the soft capsule contains one or more active compounds dissolved or suspended in a suitable liquid. Suitable liquids include, for example, one or more fatty oils, liquid paraffin, or liquid polyethylene glycol. In addition, stabilizers are optionally added.

In another embodiment, a therapeutically effective amount of one or more of the compounds described herein is formulated for buccal or sublingual administration. Formulations suitable for buccal or sublingual administration include, for example, tablets, lozenges or gels. In another embodiment, the compounds described herein are formulated for parenteral use, including formulations suitable for bolus injection or continuous infusion. In a specific embodiment, injectable formulations are provided in unit dosage forms (e.g., ampoules) or in multi-dose containers. Preservatives are optionally added to the injection formulation. In another embodiment, the pharmaceutical compositions of the compounds (i. E., The cyclohexenone compounds described herein) are formulated as sterile suspensions, solutions or emulsions in oily or aqueous vehicles in a form suitable for parenteral injection. The parenteral injection formulation optionally contains a formulation agent, such as a suspending, stabilizing and / or dispersing agent. In a specific embodiment, the pharmaceutical combination for parenteral administration comprises an aqueous solution of the active compound in water-soluble form. In a further embodiment, suspensions of the active compounds are prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles for use in the pharmaceutical compositions described herein include, for example, fatty acids such as sesame oil or synthetic fatty acid esters such as ethyl oleate or triglyceride, or liposomes. In certain specific embodiments, the aqueous injection suspension contains a component that increases the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. Optionally, the suspension contains suitable stabilizers, or agents which make it possible to increase the solubility of the compounds and make highly concentrated solutions. Alternatively, in another embodiment, the active ingredient is in powder form for constitution with a suitable vehicle, e. G. Sterile pyrogen-free water, before use.

In one embodiment, the compound (i. E., The cyclohexenone compound described herein) is prepared as a parenteral injectable solution as described herein or as known in the art, and is administered by an automatic syringe. Automatic injectors such as those disclosed in U.S. Patent Nos. 4,031,893, 5,358,489, 5,540,664, 5,665,071, 5,695,472, and WO 2005/087297, each of which is incorporated herein by reference, are known. Generally, all automatic syringes contain a volume of solution containing the compound being injected (i. E., The cyclohexenone compound described herein). Generally, an automatic syringe includes a container for holding a solution, in fluid communication with a needle for delivering the drug, and a mechanism for automatically positioning the needle and inserting the needle into the patient and delivering the dose to the patient . Exemplary syringes provide solutions at approximate concentrations of 0.5 mg to 50 mg of the compound per mL of solution (i. E., The cyclohexenone compounds described herein), about 0.3 mL, 0.6 mL, 1.0 mL, or other suitable volume. Each syringe can deliver only a single dose of the compound.

In another embodiment, the compound (i. E., The cyclohexenone compound described herein) is administered locally. The compounds described herein are formulated in a variety of topically administrable compositions such as solutions, suspensions, lotions, gels, pastes, medicated sticks, chestnuts, creams or ointments. Such pharmaceutical compositions optionally contain a solubilizer, a stabilizer, a resilience enhancer, a buffer and a preservative.

In another embodiment, the compound (i. E., The cyclohexenone compound described herein) is administered in a conventional manner, such as an enema, a rectal gel, a rectal foam, a rectal aerosol, a suppository, a jelly suppository or a cocoa butter or other glyceride Suppository bases and synthetic polymers such as polyvinylpyrrolidone, PEG, and the like. In the suppository form of the composition, the low temperature molten wax, such as, but not limited to, a mixture of fatty acid glycerides, optionally with cocoa butter, is first melted.

In certain embodiments, the pharmaceutical compositions are formulated in any conventional manner, using one or more physiologically acceptable carriers, including excipients and excipients that may enable the active compounds to be processed into pharmaceutical preparations. Suitable formulations will depend on the chosen route of administration. Any pharmaceutically acceptable technique, carrier and excipient may be used as is suitable and optionally as is known in the art. A pharmaceutical composition comprising a compound (i. E., A cyclohexenone compound as described herein) can be formulated by conventional mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, Can be prepared in a conventional manner.

The pharmaceutical compositions comprise one or more pharmaceutically acceptable carriers, diluents or excipients and one or more of the compounds described herein as the active ingredient (i. E., The cyclohexenone compounds described herein). The active ingredient is in the free acid or free base form, or a pharmaceutically acceptable salt form. In addition, the methods and pharmaceutical compositions described herein include the use of active forms of these compounds with crystalline forms (also known as polymorphs) and the same type of activity. All tautomers of the compounds described herein are included within the scope of the compounds provided herein. In addition, the compounds described herein encompass solvated as well as unsolvated forms with pharmaceutically acceptable solvents such as water, ethanol, and the like. Solvated forms of the compounds provided herein are also contemplated as disclosed herein. In addition, the pharmaceutical composition may optionally comprise other medicaments or medicaments, carriers; Preservatives, stabilizers, wetting or emulsifying agents, solubility enhancing agents, salts for controlling osmotic pressure, buffers and / or other pharmaceutically-acceptable ingredients.

A method of preparing a composition comprising a compound described herein comprises combining a compound with one or more inert pharmaceutically acceptable excipients or carriers to form a solid, semi-solid or liquid. Solid compositions include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets, and suppositories. The liquid composition may be a solution in which the compound is dissolved, an emulsion containing the compound; Or a solution comprising liposomes, micelles or nanoparticles comprising the compounds disclosed herein. Semi-solid compositions include, but are not limited to, gels, suspensions, and creams. Forms of the pharmaceutical compositions described herein include liquid solutions or suspensions, or solid forms, or emulsions, suitable for solutions or suspensions in liquids prior to use. These compositions also optionally contain minor amounts of non-toxic auxiliary components such as wetting or emulsifying agents, pH buffering agents and the like.

In some embodiments, the pharmaceutical compositions comprising at least the compound (i. E., The cyclohexenone compounds described herein) illustratively take the form of a liquid in which the drug is present as a solution, suspension or both. Typically, when the composition is administered as a solution or suspension, a first portion of the agent is present in the solution and a second portion of the agent is present in the suspension in the liquid matrix in particulate form. In some embodiments, the liquid composition comprises a gel formulation. In another embodiment, the liquid composition is aqueous.

In certain embodiments, the pharmaceutical aqueous suspension comprises one or more polymers as a suspending agent. Polymers include cellulosic polymers, for example, water-soluble polymers such as hydroxypropylmethylcellulose, and water-insoluble polymers such as cross-linked carboxyl-containing polymers. Specific pharmaceutical compositions described herein include, for example, carboxymethylcellulose, carbomer (acrylic acid polymer), poly (methyl methacrylate), polyacrylamide, polycarbophil, acrylic acid / butyl acrylate copolymer, sodium alginate and dextran &Lt; / RTI &gt; and mucoadhesive polymers selected from polyvinylpyrrolidone.

The pharmaceutical compositions also optionally include solubilizing agents to aid in the solubility of the compounds (i. E., The cyclohexenone compounds described herein). The term "solubilizing agent" generally includes a micelle solution of a drug or a drug that forms a true solution. Certain non-ionic surfactants that are acceptable, such as polysorbate 80, are useful as solubilizers, as are glycols, polyglycols such as polyethylene glycol 400 and glycol ethers, which are acceptable in ophthalmic applications.

The pharmaceutical compositions may also optionally comprise an acid such as acetic acid, boric acid, citric acid, lactic acid, phosphoric acid and hydrochloric acid; Bases such as sodium hydroxide, sodium phosphate, sodium borate, sodium citrate, sodium acetate, sodium lactate and tris-hydroxymethylaminomethane; And one or more pH adjusting agents or buffers, including buffering agents such as citrate / dextrose, sodium bicarbonate, and ammonium chloride. Such acids, bases and buffers are included in amounts necessary to maintain the pH of the composition within an acceptable range.

In addition, the pharmaceutical compositions optionally comprise one or more salts in the amounts necessary to bring the osmotic pressure of the composition to an acceptable range. Such salts include those with sodium, potassium or ammonium cations and chloride, citrate, ascorbate, borate, phosphate, bicarbonate, sulfate, thiosulfate or bisulfite anion; Suitable salts include sodium chloride, potassium chloride, sodium thiosulfate, sodium bisulfite, and ammonium sulfate.

Other pharmaceutical compositions optionally comprise one or more preservatives to inhibit microbial activity. Suitable preservatives include mercury-containing components such as &lt; RTI ID = 0.0 &gt; Stabilized chlorine dioxide; And quaternary ammonium compounds such as benzalkonium chloride, cetyltrimethylammonium bromide, and cetylpyridinium chloride.

Other pharmaceutical compositions include one or more surfactants to enhance physical stability or for other purposes. Suitable non-ionic surfactants include polyoxyethylene fatty acid glycerides and vegetable oils such as polyoxyethylene (60) hydrogenated castor oil; And polyoxyethylene alkyl ethers and alkyl phenyl ethers such as octoxynol 10, octoxynol 40.

Other pharmaceutical compositions may include one or more antioxidants to improve chemical stability if desired. Suitable antioxidants include, for example, ascorbic acid and sodium metabisulfite.

In certain embodiments, the pharmaceutical aqueous suspension composition is packaged in a single dose container that can not be closed again. Alternatively, a multi-dose container that can be closed again is used, in which case it is typical to include a preservative in the composition.

In other embodiments, other delivery systems for hydrophobic pharmaceutical compositions are used. Liposomes and emulsions are examples of delivery vehicles or carriers herein. In certain embodiments, organic solvents such as N-methyl pyrrolidone are also used. In a further embodiment, a sustained release system such as a semipermeable matrix of a solid hydrophobic polymer containing a therapeutic agent is used to deliver the compounds described herein. A variety of sustained release materials are useful herein. In some embodiments, the sustained-release capsules release the compound for a period of from several hours to 24 hours. Depending on the chemical nature and biological stability of the therapeutic agent, additional strategies for protein stabilization can be used.

In certain embodiments, the formulations described herein include one or more antioxidants, metal chelating agents, thiol-containing compounds, and / or other common stabilizers. (B) about 0.1% to about 1% w / v methionine; (c) about 0.1% to about 2% w of monothioglycerol; (e) about 0.01% to about 2% w / v of ascorbic acid, (f) 0.003% to about 0.02% w / v of polysorbate 80, (I) heparin, (j) dextran sulfate, (k) cyclodextrin, (l) pentanoic polysulfate and other heliophenides, ( m) divalent cations such as magnesium and zinc, or (n) combinations thereof.

General Considerations for Concurrent Treatment

In general, the compositions described herein, and the concurrent therapy and other agents in the embodiments described herein need not be administered with the same pharmaceutical composition, and in some embodiments, administered by different routes due to different physical and chemical characteristics. In some embodiments, the initial administration is performed according to an established protocol, and then, based on the observed effect, the skilled physician changes the dosage, mode of administration, and frequency of administration.

In some embodiments, the therapeutic effect dose is varied when the drug is used in a concurrent treatment. Concurrent therapy additionally includes periodic treatments that start and stop at various points to aid in patient clinical management. In the case of the concurrent therapy described herein, the dose of the co-administered compound will depend on the type of concurrent medication used, the particular medication used, the disease being treated, the disease or condition, and the like.

In some embodiments, the method of administration for treating, preventing, or alleviating the disease (s) to alleviate is believed to vary according to various factors. These factors include the disease to which the individual is afflicted, and the age, weight, sex, diet, and medical condition of the individual. Therefore, in another embodiment, the method of administration actually employed varies widely and therefore deviates from the administration method described herein.

Example

Example  1. Exemplary Cyclohexenone  Preparation of compounds

Through a specific herbal cleansing procedure: The following procedure is for illustrative purposes only. Other purification methods can also be used.

100 g of hyphae, fruiting bodies or mixtures of Antrodia camphorata were placed in a flask. Appropriate amounts of water and alcohol (70-100% alcohol solution) were added to the flask and stirred at 20-25 C for over 1 hour. The solution was filtered through a filter and 0.45 mu m membrane, and the filtrate was collected as an extract.

High performance liquid chromatography (HPLC) analysis was performed on the filtrate of Antrodia camphorata. Separation was carried out on an RP18 column and the mobile phase consisted of methanol (A) and 0.3% acetic acid (B), gradient conditions were 95% -20% B for 0-10 minutes, 20% -10% B for 10-20 minutes, 10 to 10% B for 20 to 35 minutes, 10 to 95% B for 35 to 40 minutes (flow rate: 1 ml / min). The column effluent was monitored with a UV-visible light detector.

The fractions collected at 25-30 minutes were collected and concentrated to give the pale yellow brown liquid product, 4-hydroxy-2, 3-dimethoxy-6-methyl-5- (3,7,11-trimethyldodeca- 6,10-trienyl) cyclohex-2-enone (Compound 1). The analysis of Compound 1 showed a molecular formula of C ₂₄ H ₃₈ O ₄ , a molecular weight of 390 and a melting point of 48 to 52 ° C. NMR spectra were determined by ¹ H-NMR (CDl ₃ )? (Ppm) = 1.51, 1.67, 1.71, 1.75, 1.94, 2.03, 2.07, 2.22, 2.25, 3.68, 4.05, 5.07 and 5.14; _{^{13 C-NMR (CDCl 3)}} δ (ppm) = 12.31, 16.1, 16.12, 17.67, 25.67, 26.44, 26.74, 27.00, 39.71, 39.81, 40.27, 43.34, 59.22, 60.59, 120.97, 123.84, 124.30, 131.32, 135.35 , 135.92, 138.05, 160.45 and 197.12.

Compound 1: 4-Hydroxy-2,3-dimethoxy-6-methyl-5- (3,7,11-trimethyldodeca-2,6,10-trienyl) cyclohex-

R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl; Which is optionally substituted by one or more substituents selected from C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl and C ₁ -C ₈ haloalkyl, Other compounds which are aryl have also been obtained.

Alternatively, exemplary compounds may be prepared by synthesis. See, for example, U.S. Patent No. 9365481.

의 구조를 갖는 다른 사이클로헥세논 화합물은 안트로디아 캄포라타로부터 단리되거나, 또는 적합한 출발 물질로부터 합성에 의해 또는 반-합성에 의해 제조된다. 당 업자는 이러한 합성에 적절한 조건을 용이하게 활용한다.Similarly,

Are isolated from the antrodia camphorata or are prepared by synthesis or by semi-synthesis from suitable starting materials. The person skilled in the art will readily utilize appropriate conditions for such synthesis.

Example  2. AsPC -1 cell culture system and severe combined immunodeficiency mouse xenograft tumor model in human Clinical Chemotherapy for Pancreatic Cancer  Study of Drugs and Compounds 1

To assess the efficacy of a combination therapy comprising an anticancer agent (e. G., Any of the clinically used drugs) for the treatment of pancreatic cancer and the exemplary Compound 1, the currently selected clinical chemotherapeutic drug In vitro and in vivo xenograft studies were performed.

For in vitro studies, Compound 1 and each of the six selected clinical chemotherapy drugs were used in human pancreatic cancer AsPC-1 and / or CaPan-2, PANC-1 in a cell culture system.

In a xenotransplantation mouse model, gemcitabine and / or paclitaxel were selected with oral administration of Compound 1 once a day for a total of 28 doses in the human pancreatic cancer AsPC-1 xenograft model, Lt; / RTI &gt;

In vitro  exam:

Human pancreatic cancer cell lines AsPC-1, PANC-1, and CaPan-2 were inoculated at 3 × 10 ⁴ cells per well and cultured in 48-well cell culture dishes (growth area per well: 0.95 cm ² ). After 16 hours, the cells were treated with compound 1 alone or with compound 1, respectively. After 48 or 72 hours of treatment, methylthiazole tetrazolium (MTT) assay (Supplementary Method-1) was performed to determine the effect of treatment on cell viability during proliferation.

In vivo  exam:

Human pancreatic cancer cell line AsPC-1 was cultured and cells were subcutaneously injected into the flank of 60 mice. Three days later, mice were able to feel the tumor. When tumors reached about 100 to 200 mg (mm &lt; ³ &gt;), a total of 51 tumor-bearing mice were randomly selected and sorted into one of six groups. Mice treated with vehicle (corn oil) were used for negative control. Compound 1 in combination with gemcitabine or paclitaxel was administered intravenously once per day for four weeks and / or intravenously two times per week. Tumor measurements were recorded using a portable caliper and a Visual Sonic Vevo 2100 imaging system. Individual mice were weighed once a week during dosing. Mice are monitored daily for signs of toxicity and morbidity; Abnormal results were recorded. The study was terminated four weeks after initiation of drug administration.

The mice were euthanized when individual mice had tumors over 2,000 mg (cm &lt; ³ &gt;) at one time of measurement. When the tumor interferes with the ability of the mouse to eat, drink or walk, or when the original weight of the mouse is reduced by more than 20%, the mouse is in a dull state. Individual groups were euthanized when tumors reached an average target size of 1,500 mg per group over two consecutive measurements. The researcher should know the unexpected euthanasia. The study was terminated four weeks after initiation of drug administration.

Test system:

A. Cell line:

Human pancreatic cancer AsPC-1 (ATCC catalog number CRL-1682).

Human pancreatic cancer PANC-1 (ATCC catalog number CRL-1469).

Human pancreatic cancer CaPan-2 (ATCC catalog number HTB-80).

B. Cell preparation

1. Remove and discard the culture medium.

2. The cell layer is simply rinsed with 5 mL HBSS. Gently shake and remove HBSS.

3. Add 5 mL trypsin-EDTA to a 15 cm ² polystyrene dish. In a humidified incubator with a 95% air / 5% CO ₂ atmosphere and incubated at 37 ℃ 5 minutes.

4. Add 5 mL of complete growth medium (DMEM medium containing 10% FBS) and inhale the cells by gently pipetting.

5. Count the viable cells with a trypan blue exclusion assay (supplementation method-2) as a hemocytometer.

6. For in vitro studies, cells are inoculated with 3 x 10 ⁴ cells per well and cultured for 16 h until treated in a 48-well cell culture dish.

7. Cultivate the culture at 37 ° C.

C. Animal Species / Strain:

a. Nude mice, BALB / cAnN.Cg- Foxnl ^nu / CrlNarl.

b. Female, athymic nude mice (nu / nu) 4 to 6 weeks old.

c. Weight: 15-20 g on the day of tumor cell inoculation.

D. Human pancreatic cancer xenograft model:

1. To form a subcutaneous xenograft model, athymic female nude mice (4-6 weeks old) were inoculated with 0.1% serum-free DMEM medium containing 20% Matrigel [BD Biosciences, Bedford, MD] an mL 1 × 10 ⁷ cells of the gae AsPC-1 were inoculated subcutaneously.

2. Tumor-bearing animals were divided into the following six groups as shown in Table 1:

3. The vigor and physical condition of the mice should be monitored daily and weight determination and tumor mass measurements should be performed once a week. Take a picture of the tumor image by digital camera and Visual Sonic Bebo 2100 imaging system. The tumor mass was determined by measuring the two orthogonal diameters of the grafts with a caliper and the tumor mass was determined by the formula (4/3) x (? A b ² ) (where "a" Short diameter).

E. Test Products:

Clinical chemotherapy drugs were purchased from Sigma-Aldrich Co. LLC and were prepared in the following manner:

-Allotinib (100 mM in total concentration DMSO, Sigma catalog # E4997).

- 5-fluorouracil (parent concentration 100 mM, Sigma catalog # F6627).

- gemcitabine (100 mM in total concentration DMSO, Sigma catalog # G6423).

- Irinotecan (100 mM in parent DMSO, Sigma Catalog # I1406).

- oxaliplatin (50mM in parent DMSO, Sigma catalog # O9512).

- Paclitaxel (10 mM in parent DMSO, Sigma catalog # T7402).

For in vivo testing, gemcitabine (G) was purchased from Eli Lilly (Indianapolis, Ind.) And paclitaxel (P) was purchased from Sigma-Aldrich.

F. In vivo  Experimental Design:

a. Cytotoxicity (TC ₅₀ ) of the test product : AsPC-1 was inoculated with 3 × 10 ⁴ cells per well (counted by a cell counter, LUNA ™ automated cell counter or blood count plate), and 48 well cells And cultured in a culture dish. Cells were treated with 400 [mu] M to 800 [mu] M single product with continuous dilution to 0 [mu] M. MTT assay was performed 48 or 72 hours after treatment to determine the effect of treatment on the survival rate of AsPC-1 cells during proliferation. The absorbance at 595 nm was measured using a microplate reader (BioTEK, Synergy ™ H1). Product sensitivity curves and TC ₅₀ values were calculated using GraphPad Prism 4.0 software.

b. Concurrent therapy of Compound 1 and chemotherapy drugs: This study also involves the cytotoxicity of Compound 1 and selected chemotherapeutic drugs to determine the efficacy of the combination. Compound 1 was combined with paclitaxel, gemcitabine, 5-FU, oxaliplatin, allotinib and irinotecan, respectively. 72 hours after treatment, MTT analysis was performed to determine the effect of treatment on the survival rate of AsPC-1 cells during proliferation. The absorbance at 595 nm was measured using a Microwell plate reader.

c. Assessment of Drug Interactions: MTT assays were also performed to test the effect of drug combinations on survival rates of AsPC-1, PANC-1 and CaPan-2 cell lines. Chou and Talalay combination index (CI) based on the median effect principle. The isobologram method [Chu and Talalay, 1984; Chang et al., 2000], synergistic or antagonistic effects on the combined drug effects were calculated. The dose-effect curves of each drug (individually and in combination) in the sub-doses of diluted concentrations were plotted using median effect equations and plots (Chu, 1991) and CI equations and plots (Chu et al., 1994) . Using computer software CompuSyn [Chu and Martin, 2005], CI values and isovolumograms at different effects and dose levels were automatically generated. In this way, additive, ascending, or antagonistic effects are indicated by CI values of 1, < 1 and > 1, respectively.

G. In vivo experiment design:

a. Cell injection: 1 x 10 ⁷ living human pancreatic cancer AsPC-1 cells were subcutaneously injected into the right lateral chest of mice. A total of 60 mice were injected and about 36 tumor-bearing mice were used in this study.

b. Animals Selection and Mice Assignment: The study included 6 groups, each containing 8 or 9 mice each. When tumors reached a target size of about 100 to 200 mg (mm ³ ), 54 tumor-bearing mice were randomly selected and divided into 6 groups. Group 1 was a solvent-treated negative control, and received treatment-1. Groups 2, 3, 4, 5 and 6 included mice receiving treatment-2, treatment-3, treatment-4, treatment-5 and treatment-6, respectively. Table 1 lists the test product concentration, solvent product and dosing schedule.

c. Administration formulations:

c-1: Compound 1 (A)

i. Weigh the appropriate amount of Compound 1 and place it in a 50 mL tube.

ii. Add the appropriate amount of corn oil (Sigma-Aldrich) to the 50 mL tube. Mix the formulation with the vortex until it is completely homogeneous. The formulation is placed into a disposable syringe and transferred to a 4 mL brown tube (3.2 mL / tube).

iii. Store the suspension under freezing conditions (-20 ° C) until use.

iv. The formulation is completely thawed at 37 DEG C in a water bath on the intended use day.

c-2: Gemcitabine (G)

i. Commercially available gemcitabine was stored as a 50 mg / mL solution in sterile PBS at -20 &lt; 0 &gt; C [Ito et al., 2006; Awasthi et al., 2013].

ii. Before animal studies, an appropriate amount of gemcitabine was weighed into 4 mL tubes.

iii. Appropriate volumes of Vehicle-I (50% ethanol and 50% Tween-80 (v / w)) were added to the 4 mL tubes and shaken until the material was completely dissolved.

iv. Appropriate volumes of physiological saline were added to the tubes in 4 mL and the tubes were shaken sideways to completely homogenize the solution.

c-3: paclitaxel (P)

i. Commercial paclitaxel was dissolved in 100% ethanol to a final concentration of 10 mg / mL (Shi et al., 2000; Chang et al., 2006; Awashushi et al., 2013].

ii. Prior to animal study, the appropriate amount of paclitaxel was weighed into 4 mL tubes.

iii. The vehicle -I ^b an appropriate volume of 4mL are added to the tube, and shook until the substance is completely dissolved.

iv. Appropriate volumes of physiological saline were added to the tubes in 4 mL and the tubes were shaken sideways to completely homogenize the solution.

Group number Treatment Label Dose / mouse / day or number of times Route of administration Dosage volume / mouse frequency One Treatment -1 Solvent (corn oil) oral- 100 μL for oral administration, 20 to 30 μL for I.V. Once per day for oral administration for 4 weeks, twice per week for IV 2 Therapy-2 A (120 mg / kg) Oral (A) 3 Treatment -3 A (120 mg / kg)
G (100 mg / kg) Oral (A) / IV (G) 4 Treatment -4 A (120 mg / kg)
P (5 mg / kg) Oral (A) / IV (P) 5 Treatment -5 G (100 mg / kg)
P (5 mg / kg) IV (G & P) 6 Treatment -6 G (100 mg / kg)
P (30 mg)
A (120 mg / kg) Oral (A) / IV (G & P)

Table 1. Treatment groups of AsPC-1 xenografts

Observation and review of in vivo systems A. Clinical observations: Clinical observations of each study animal were performed at least once daily (including weekends and holidays) for morbidity, mortality and signs of toxicity of the test product. The morbidity rate includes signs of illness such as, but not limited to, dehydration, dehydration, lethargy, bent posture, flattened appearance, dyspnea and urinary or fecal contamination. Record any abnormal observations.

B. Animal weights: Individual animal weights were measured once per week before / before dosing. First, body weight was recorded before cell injection and weight was recorded continuously throughout the study.

C. Tumor measurement: The injection site of each animal was monitored once a week for tumor growth indications. Throughout the study, the length (a) and width (b) of any tumors generated were measured in mm using a vernier caliper (where a is the longer of the two dimensions) . The applicable tumor size (mm or cm) was calculated by using the equation M = (4/3) x (pi x a x b ² ) associated with the circular ellipsoid, and these data were recorded.

D. Assessment of drug synergistic effects by tumor growth inhibition (TGI) and tumor suppression ratio (TIR): The tumor growth inhibition (TGI) volume of each group was calculated according to the following equation:

Tumor growth inhibition ( % ) = (1- [T-T ₀ ] / [C-C ₀ ] × 100

T and T ₀ are the mean tumor volume on the treatment end date and on the first day of treatment in the experimental group, respectively. For the control group, C and C ₀ are the mean tumor volume at study end date and onset day, respectively.

In the case of TIR, after 4 weeks of treatment, the mice were killed, the tumors were carefully excised, and any residual blood was washed with PBS and weighed. The tumor suppression ratio was calculated by the following equation:

Tumor suppression ratio ( % ) = [( W _Control - W _cure ) / W _Control ] × 100

W and W _{therapy for quiet} a mean tumor weight of each mouse and treatment for a quiet mouse.

D. Hematological parameters: WBC, RBC, hemoglobin and concentration, hematocrit, mean hemoglobin and hemoglobin concentration, platelet distribution width, platelet count, neutrophil, lymphocyte, mononuclear cell, eosinophil To analyze hematological parameters, cardiac blood was collected in EDTA. This was determined using a blood counter (Abbott Cell-Dyn 3700, IL).

Statistical analysis: All data are presented as mean ± standard error of the mean (SEM). The significance between groups was determined using one-way ANOVA. To compare more than two groups more than once, ANOVA analysis was used followed by post-test using Bonferroni correction. A value of P < 0.05 was considered significant.

A day of xenotransplantation studies Schedule  example:

result:

In vitro cytotoxicity: To evaluate the concurrent therapy efficacy of the exemplary Compound 1, the 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) The survival rate of the treated AsCp-1 cell line was measured. The toxic concentration (TC ₅₀ ) of Compound 1 (i.e., anthroquinol) and the chemotherapeutic drug against the AsPC-1 cell line was examined. Figures 1 to 7 and Table 2 show the cytotoxicity of Compound 1 and the chemotherapeutic drug against the AsPC-1 cell line. Based on these results, the concurrent treatment of Compound 1 and the chemotherapeutic drug against the AsPC-1 cell viability according to the drug concentration in Table 3 was confirmed. AsPC-1 cells were individually cultured with Compound 1 or each drug combination for 48 hours and 72 hours. In Figures 8 and 9, the MTT assay of each chemotherapeutic drug and their combination was determined for survival rates of AsPC-I cells incubated with compound 1, 48 hours and 72 hours, respectively. AsPC-1 cells treated with Compound 1 or chemotherapy alone exhibited 75-80% survival after 48 or 72 hours of culture. Based on these results, the concurrent therapy with the selected chemotherapeutic drug and Compound 1 has a synergistic effect and can modulate the AsPC-1 cell survival rate. In addition, these results also indicate that the combination of the tested clinical drug and Compound 1 has a more extreme effect on AsPC-1 human pancreatic cancer cells than the corresponding single drug treatment.

drug TC ₅₀ (μM) ^a 48 hours 72 hours Anthroquinol 58.09 24.04 Allotinib > 800 > 800 5-FU > 800 > 800 Gemcitabine > 800 > 800 Irinotecan 221.3 68.34 Oxaliplatin 654.7 57.91 Paclitaxel 7.054 3.719 ^a TC ₅₀ : Concentration of compound with cell viability reduced to 50%

Table 2. Effect of anticancer drugs on cytotoxicity at 48 hours and 72 hours (Antroquinol = Compound 1)

The combination of paclitaxel and / or gemcitabine and compound 1 with synergistic effect on pancreatic cancer was evaluated. The combination was tested through an intermediate effect analysis using three pancreatic cancer cell lines AsPC-1, CaPan-2 and Panc-1.

drug Concentration (μM) 48 hours 72 hours Anthroquinol (Compound 1) 20 20 Allotinib 100 100 5-FU 100 100 Gemcitabine 100 100 Irinotecan 200 70 Oxaliplatin 200 60 Paclitaxel 7 3

Table 3. Drug concentrations used in combination for cytotoxicity of AsPC-1 cells

Chemotherapy drug Gemcitabine  And Paclitaxel  Of the concurrent therapy effect of Compound 1 Flat end

The CI of each combination combination was calculated using a median effect analysis of cholesterol and taralay (Chu and Talalay, 1984). Thus, the CI value was used to determine whether there was a synergistic effect between Compound 1 and gemcitabine and / or paclitaxel in this study. In the AsPC-1, CaPan-2 and Panc-1 cell lines, the combination of two or three compounds of the chemotherapeutic drugs paclitaxel and gemcitabine and Compound 1 span the entire dosage range (see, e.g., Figures 10 and 11) (CI < 1, Figure 11). These results showed that the second or third currently used chemotherapeutic drug and the exemplary compound (i.e., Compound 1) potently reverse pancreatic cancer growth in the combination of three compounds in particular.

AsPC -1 for subcutaneous xenograft model In vivo  Antitumor activity

The anticancer activity of the concurrent therapy including the exemplary Compound 1 was evaluated in nude mice containing AsPC-1 tumor. AsPC-1 cells were injected subcutaneously into mice 7 days prior to initiation of treatment (Table 1). These animals were then given various drug combinations daily for 28 days. On day 28, all test mice were killed and the corresponding tumors excised for further study. 13 and 14, after the experiment, mice treated with a combination of Compound 1 and gemcitabine (T3) or a combination of Compound 1, gemcitabine and paclitaxel (T6), together with slow growth of the tumor volume Showed very significant tumor regression. Mice in the other four treatment groups exhibited intact tumor growth compared to the groups T3 and T6. The tumor growth inhibition rate was significantly increased in the combination of compound 1 with gemcitabine (T3) or with compound 1, gemcitabine and paclitaxel (T6) compared to the group (Fig. 16, T2, T4 and T5) (Fig.

Figure 17 shows the treatment results provided by an ultrasonic image of an AsPC-1 pancreatic cancer xenograft during the treatment period. A photograph of the tumor from dead mice is provided in Figure 18A. It can be seen that the tumor volume of mice treated with compound 1 and gemcitabine is significantly smaller than that treated with other combinations. Specifically, Figure 18B shows that tumor weights of mice treated with compound 1 and gemcitabine are much smaller than those of the other groups. These results demonstrate that the concurrent therapies described herein all have predominant tumor growth inhibiting efficacy. The tumor inhibition ratio (TRI) of each group was also calculated based on the final tumor weight (Figure 19). Tumor growth was significantly inhibited in the group of compound 1 + gemcitabine (Fig. 19, T3) and compound 1 + the other two drugs (Fig. 19, T6).

(TCR), tumor growth inhibition (TGI), and tumor suppression ratio (TCR) of the tumor volume in order to confirm that the therapeutic efficacy of the exemplary compound 1 based concurrent therapy is much better than that of the single chemotherapeutic drug TIR), the synergistic effect of the selected drug combination was evaluated. In AsPC-1 xerografts, monotherapy of Compound 1 was able to inhibit tumor growth with a treatment-to-control ratio (TCR) of 0.55. The combination of Compound 1 and gemcitabine (Table 4, Ge + Aq) produced a dramatically improved anticancer activity compared to other single or two-compound combination therapies (TCR 0.36; Table 4; On the other hand, the combination of the three drugs including Compound 1 (Table 4, Px + Ge + Aq) also produced stronger anticancer activities than single or other parallel therapies (TCR 0.46; Table 4; Based on these results, compound 1 in combination with gemcitabine or additional paclitaxel significantly enhanced its anticancer activity.

In addition, no significant weight loss was observed in the tumor-bearing mice treated with the various combinations, indicating that the side effects of Compound 1 with the other two chemotherapy drugs for tumor therapy in the doses used (Figure 12) were negligible . Similarly, Compound 1 and its combination did not affect the number of blood cells other than blood glucose in AsPC-1 xenografts (Table 5). Based on these data, it is proposed that Compound 1 and / or combinations thereof have a low safety risk when administered orally.

Treatment group ^# T1 T2 T3 T4 T5 T6 Tumor weight 0.42 ± 0.11 ^a 0.21 ± 0.07 ^b 0.11 0.05 ^b 0.22 ± 0.16 ^b 0.31 0.21 ^b 0.24 0.09 ^{b #} Corn oil (T1), compound 1 (T2), the compound combination of 1 and gemcitabine (T3), the combination of Compound 1 and paclitaxel (T4), the combination of gemcitabine and paclitaxel (T5) and Compound 1, gemcitabine and Six treatment groups of mice treated with the combination of paclitaxel (T6).
- Weight of the weight is mean ± SEM. Values in the same row with different subscripts (ab) are significant at P < 0.05.

Table 4. Tumor weight of tumor excised at death from subcutaneous AsPC-1 pancreatic cancer xenograft-bearing male nude mice after 28 days of treatment

xenograft cure Reaction, TCR TGI (%) TIR (%) AsPC-1 Control 1.00 - - Aq 0.55 51 45 Ge + Aq 0.36 73 64 Px + Aq 0.60 49 40 Px + Ge 0.67 40 33 Px + Ge + Aq 0.46 65 54 Abbreviations: Aq, compound 1; Px, paclitaxel; Ge, gemcitabine; TCR, Tumor volume of treatment versus control ratio; TGI, tumor growth volume inhibition; TIR, tumor weight suppression ratio

Table 5. Synergistic anticancer activity by combination of compound 1 (Aq), paclitaxel (Px) and gemcitabine (Ge) in AsPC-1 xenograft

Table 6. Hematological values of each treatment group in AsPC-1 xenograft study

As shown in the results of the study, exemplary compounds 1 and a composition comprising a prospective clinical chemotherapeutic drug such as paclitaxel and gemcitabine clearly demonstrate the unexpected advantage of using against pancreatic cancer. Cell survival studies suggest that treatment with this combination produces a much higher inhibitory activity than Compound 1 or other chemotherapeutic agents alone, suggesting synergistic and / or additive effects. This effect was confirmed by the combination of compound 1 and gemcitabine alone, and in combination with additional paclitaxel, which represents an excellent inhibition of tumor growth in mice bearing tumors.

Example  3. Max Medicine  Dose ( MTD ), And the first-line metastasis from pancreatic cancer Nave + Safety of Compound 1 in combination with paclitaxel + gemcitabine / Tolerance , Pharmacokinetics , Pharmacodynamics  And for assessing temporary efficacy 1 phase  Research

INTRODUCTION : In 2011, the multidrug combination of leucovorin, fluorouracil, irinotecan and oxaliplatin (FOLFIRINOX) was found to provide an increased median survival time of 4.3 months compared to gemcitabine; Considering his side effect profile, this can only be used in selected groups of advanced pancreatic cancer patients. Patients were randomly assigned to receive either polipyrinox or gemcitabine. Overall median survival (OS) was 11.1 months in the papyrus knox group compared to 6.8 months in the gemcitabine group [hazard ratio for death (HR) = 0.57; 95% confidence interval [CI], 0.45 to 0.73; p < 0.001]. The median progression-free survival (PFS) was 6.4 months in the Porphyrinox group and 3.3 months in the gemcitabine group (HR = 0.47; 95% CI, 0.37-0.59; p <0.001 for disease progression).

More recently, the combination of gemcitabine and nab-paclitaxel (a 130 nm albumin-conjugated nanoparticle formulation of paclitaxel) has been shown to increase the median survival time by 1.8 months (1 and 2 year increased OS); Side effects were moderate and included cytopenias and peripheral neuropathy. Multi-center International Phase III MPACT experiments included 861 patients with previously untreated metastatic pancreatic adenocarcinoma. Patients were randomly assigned to receive gemcitabine and nab-paclitaxel or gemcitabine monotherapy. The median OS was 8.5 months in the Nab-paclitaxel / gemcitabine group (HR = 0.72; 95% CI, 0.62-0.83; p <0.001 for death) compared to 6.7 months for the gemcitabine group. The median PFS was 5.5 months in the Nab-paclitaxel / gemcitabine group and 3.7 months in the gemcitabine group (HR = 0.69; 95% CI, 0.58-0.82, p <0.001 for disease progression).

Currently, the National Comprehensive Cancer Network Recommendation is based on patient preference and available support systems, allowing patients with an excellent performance status (ie, an Eastern Cooperative Oncology Group (ECOG) performance status (PS) of 0 or 1) Therapy combinations. These combinations include, but are not limited to, porfirinox for patients with superior / preferred concurrent mood profiles, or gemcitabine + nab-paclitaxel, nab-paclitaxel + gemcitabine, and gemcitabine + Lt; / RTI &gt; Gemcitabine alone is recommended for patients with ECOG PS 2 or patients with a simultaneous morbidity profile that excludes other therapies; Addition of capecitabine or allotinib may be provided. Thus, guidelines for selecting appropriate therapies for patients with metastatic pancreatic cancer remain ambiguous, and the most appropriate frontline therapies for these patients remain unclear in the absence of randomized trials comparing concurrent therapies .

As shown in Example 2, exemplary Compound 1 has been shown to have anticancer activity and a low toxicity profile prior to clinical in vitro and in vivo animal studies. In a separate non-clinical study using the PANC-1 human pancreatic cancer xenograft model performed with the Compound 1 in 2012, four groups of mice were treated with 30 mg / kg, 60 mg / kg, 90 mg / kg Compound 1 And vehicle control to investigate the in vivo therapeutic efficacy of Compound 1. Tumor volume and tumor weight were measured 10 days after drug treatment (19 days after tumor implantation). Treatment with 30, 60 and 90 mg / kg of Compound 1 produced effective anticancer activity with statistically significant smaller mean tumor volumes and tumor weights at all three dose levels compared to vehicle control. All three doses of anthroquinolone were tolerated in tumor-bearing mice; No significant weight loss was observed during the study.

In an earlier reported clinical trial study, anthroquinolol (i.e., Compound 1) at 50, 100, 200, 300 450 and 600 mg dose levels daily administered for 4 weeks showed no significant safety issues or DLT in this study Since it was not identified, it was generally safe and well tolerated. The data generated from the safety and pharmacokinetic (PK) profiles in this study are used to determine the maximum tolerated dose (MTD) and efficacy in a given patient population by the addition of an additional amount of anthroquinol in an individual with advanced malignancy I was able to support the study.

Research Purpose

The primary objective of this study is to determine the MTD or the maximum tolerated dose (MFD) of Compound 1 in combination with Nab + paclitaxel + gemcitabine in an individual with metastatic pancreatic cancer. Although Compound 1 has been evaluated for cancer treatment, it is known in the art that many cancer drugs are less effective in treating metastatic cancer.

Group expansion: To evaluate the anticancer activity of anthraquinolone in combination with nab + paclitaxel + gemcitabine in individuals with metastatic pancreatic cancer.

The secondary objectives of this study are as follows:

Increased dosage:

To assess the safety and tolerability of a combination of anthraquinolone and nab + paclitaxel + gemcitabine.

To characterize PK of anthraquinolone and nab + paclitaxel + gemcitabine.

To assess anthraquinone activity from normal pancreatic cancer monitoring.

To explore the preliminary anticancer activity of anthraquinolone in combination with Nab + paclitaxel + gemcitabine in individuals with metastatic pancreatic cancer.

Group expansion: To assess the safety and tolerability of MFDs in combination with anthraquinolone and nab + paclitaxel + gemcitabine; To characterize PK of anthraquinolone and nab-paclitaxel-gemcitabine; Assessing anthraquinone activity from conventional pancreatic cancer monitoring for.

Research endpoint

Primary end point

Increased dosage:

• The occurrence of DLT in the first 28-day cycle of treatment. DLT Definition [Class according to NCI CTC (Common Terminology Criteria for Adverse Event) v.4.03]

● Third grade diarrhea that resolves to grade 1 or baseline grade within 3 days of initiation of appropriate treatment, non-hematologic toxicity of grade 3 or higher, excluding nausea or vomiting

● 4-grade thrombocytopenia or leukopenia lasting more than 48 hours

● Grade 3 or more febrile leukopenia

● Grade 3 thrombocytopenia with bleeding

● Dose A sustained toxicity of more than grade 2, resulting in a delay of more than 14 days in administration.

AEs considered by investigators to be related to underlying disease, concurrent medications, or new unrelated medical treatment or treatment are not defined as DLTs.

Group expansion:

Primary endpoint: To assess the anticancer activity of anthraquinolone in combination with nab + paclitaxel + gemcitabine in individuals with metastatic pancreatic cancer.

Secondary endpoints:

Safety: Safety is assessed using AE, physical examination, laboratory observation (including clinical chemistry, hematology and urinalysis), vital signs (including blood pressure and pulse), and electrocardiogram (ECG).

Efficacy: PFS, OS, using investigative assessment according to objective response rate (ORR), duration of response (DoR), disease inhibition rate (DCR), and RECIST 1.1 (evaluation every 8 weeks).

Pharmacokinetics : The plasma concentration and PK parameters of anthraquinolone and Nab + paclitaxel + gemcitabine combination are evaluated.

Biological Indicators: To assess the anthraquinone nonol activity from the usual pancreatic cancer monitoring.

Investigation plan

Full research design and planning description

) 개체의 제일선 치료에서 나브-파클리탁셀 및 겜시타빈과 조합된 화합물 1의 단일-암(single-arm) 개방-표지 다중 중심 I상 연구이다.This is due to the fact that IV

) Is a single-arm open-label multiple center phase I study of Compound 1 in combination with nab-paclitaxel and gemcitabine in the first line therapy of the individual.

The first part of the study follows a 3 + 3 dose escalation design to determine the MTD or MFD (based on PK and capsule strength) of anthraquinolone combined with standard dosing regimens of nab-paclitaxel and gemcitabine. Two dose levels of anthraquinolone provided three times a day are planned, based on the dose increasing data and the capsule strength. The final number of individuals is based on the number of dose levels being examined and the occurrence of DLT. Up to 12 individuals can be treated in this part of the study. A Safety Monitoring Committee (SMC) will be established to review the DLT and overall safety data for this study and determine the relevance of the treatment for the dose escalation plan.

After MTD / MFD determination, registration will continue at a fixed dose of anthraquinolone (MTD or MFD). A maximum of 40 individuals, which can be evaluated, are planned for this population expansion part of the experiment.

Investigation product, dose and method of administration

● Increase the dose of Compound 1 (200-300 mg) three times a day by oral administration until there is unacceptable toxicity or disease progression without interruption criteria.

• 125 mg / m ² nave-paclitaxel and 1000 mg / m ² gemcitabine via intravenous (IV) infusion, both until the unacceptable toxicity or disease progression, on the first day of each 28- 8 &lt; / RTI &gt; and 15 days (i. E., Weekly for 1 cycle = 3 weeks, no further weeks thereafter).

DLT definition and evaluation

Dosage restriction toxicity is defined as the occurrence of any of the following toxicities that occur during the DLT evaluation period, which is determined to be related or related to the combination of the investigational medicinal product (IMP). The DLT evaluation period is the first 28-day treatment cycle. Toxicity ratings are determined using NCI CTCAE version 4.03. Dosage restriction toxicity is defined as:

● Third grade diarrhea that resolves to grade 1 or baseline grade within 3 days of initiation of appropriate treatment, non-hematologic toxicity of grade 3 or higher, excluding nausea or vomiting

● 4-grade thrombocytopenia or leukopenia lasting 248 hours

● Grade 3 febrile leukopenia

● Grade 3 thrombocytopenia with bleeding

● Dose A sustained toxicity of more than grade 2, resulting in a delay of more than 14 days in administration.

AEs considered by investigators to be related to underlying disease, concurrent medications, or new unrelated medical treatment or treatment are not defined as DLTs.

Selection of study populations

Inclusion Criteria: In order to be eligible to participate in the study, an individual must meet the following criteria:

1. Male and female objects over 18 years of age.

2. Measurable, histologically or cytologically confirmed metastatic pancreatic adenocarcinoma according to recyst 1.1.

3. Metastatic disease should be diagnosed within 6 weeks before randomization.

4. Therapeutic-naive individuals with metastatic pancreatic adenocarcinoma who have not been previously treated with systemic therapy (except for adjuvant or neoadjuvant therapy if progress has occurred beyond 6 months after final treatment or surgery; no preceding nava-paclitaxel).

5. Hemoglobin ≥ 9 g / dL

- absolute neutrophil count ≥1500 / mm ³

- Platelet count ≥100 000 / mm ³

- total bilirubin ≤1.5 × normal upper limit (ULN), except for individuals with documented Gilbert's syndrome (> 3 × ULN)

- alanine aminotransferase (ALT) and aspartate aminotransferase (AST) &lt; 2.5 x ULN; For patients with lung metastases, ALT and AST ≤ 5 × ULN

- Albumin ≥3mg / dL

- serum creatinine ≤1.5 mg / dL or Cockcroft - creatinine clearance calculated by the Gault equation ≥50 mL / min

- 0 or 1 ECOG

Including proper blood, lung and kidney function.

6. Results of negative serum pregnancy test on the day of selection and day 1 in the case of the reproductive female.

7. Use two medically acceptable and effective methods of contraception during the study (both male and female, if appropriate) and three months after the last dose of study medication, from the list below:

- Established use of oral, injectable or transplanted hormonal methods of contraception.

- Intrauterine device or intrauterine system setup

- Blocking contraceptive methods: closed caps (septum or cervix / bolt cap) with condom or sachet foam / gel / film / cream /

- Male infertility (with proper documentation of the absence of semen in the absence of ejaculation)

- True abstinence: If this is desirable and consistent with the lifestyle of a typical individual,

8. Signed ICF

9. Life expectancy over 12 weeks.

Exclusion criteria: Individuals meeting any of the following criteria can not participate in the study:

1. Islet neoplasm or locally advanced disease.

2. Within 4 weeks or within 5 half-lives of the study drug (whichever is shorter) the date of the first administration of chemotherapy, hormone therapy or immunotherapy or study medication and / or anticancer therapy that appears to be clinically relevant Continued toxicity.

3. Treatment with any drug (s) known to be potent inhibitors or inducers of cytochrome P450 (CYP) 2C19, CYP3A4, CYP2C8 and CYP2E1 within 14 days of the first administration of the study drug and during the study.

4. Other malignancies diagnosed within the past 5 years, including cervical carcinoma treated by treatment, non-melanoma skin cancer, shallow bladder cancer Ta [noninvasive tumor] and TIS [epithelial cell cancer], or surgery or radiation therapy And the serum prostate-specific antigen is within the normal range [testing conducted within the past 12 months prior to the first administration of the study drug], except for Vision 1 or 2 prostate cancer.

5. Individuals with any serious active infection (i. E., Requiring intravenous antibiotics, antifungal or antiviral agents).

6. Individuals with human immunodeficiency virus, active hepatitis B, or active hepatitis C.

7. Any entity with any other fatal disease or organ system dysfunction that, in the opinion of the investigator, sacrifices the safety of the individual or interferes with the safety assessment of the study drug.

8. Known or suspected drug abuse or alcohol abuse.

9. Failure to comply with ongoing or active infection, congestive heart failure, uncontrolled hypertension, unstable angina pectoris, cardiac arrhythmia, interstitial lung disease, or study requirements, or significantly increase the risk of developing AE from research therapy Or mental illness / social conditions that jeopardize the patient's ability to provide printed and publicized agreements.

10. Recent acute gastrointestinal disorders that do not swallow oral medications or diarrhea, such as Crohn's disease, malabsorption, or diarrhea with CTCAE grade 2 or higher of any etiologic factor on the baseline.

11. A pregnant or lactating woman, or a male or female individual capable of breeding without using effective methods of contraception.

Withdrawal of an individual: An individual may withdraw his / her consent to participate in the study, and the investigator may withdraw the individual without prejudice. If any of the conditions listed below occur, the subject may withdraw from the study or study therapy:

1. Documented disease progression according to response criteria.

2. Unacceptable toxicity.

3. The entity decides to withdraw the agreed consensus.

4. The investigator considers that an individual can not remain physically and / or mentally in the study any longer.

Alternative Procedure: During the dose increasing period, individuals who did not experience DLT in the first cycle and meet any of the following should be replaced with new individuals:

• Do not complete all scheduled experimental treatment doses within the first 28 day cycle.

• All planned experimental treatment dosing was completed without delay / delay, but 90% of the planned total dose of drug was not administered within the first cycle.

Inspector guidelines for when to terminate the study treatment plan: Individuals may withdraw their consent to participate in the study, and the investigator may withdraw from the subject at any time. An individual is withdrawn from research or study therapy if he or she meets any of the following conditions:

1. No follow-up action.

2. An arbitrary AE or medical condition where the investigator or sponsor determines that the safety of the individual may be compromised if the individual continues to study.

3. Trying to become pregnant or pregnant.

4. In the opinion of the investigator or the sponsor, the non-conformity of the entity justifying the withdrawal from the study medication (eg refusing to keep the planned visit).

5. Initiation of other chemotherapeutic regimens, including other research drugs.

6. Investigator's determination that confirmed progressive disease and study therapy are no longer beneficial to the individual.

7. The sponsor ends the study. The reasons for the termination of the study may include, but are not limited to: the occurrence or severity of AE in this or other studies indicates that this would be a health hazard to the individual; Unsatisfactory registration of the object.

Follow-up of an object that has been prematurely terminated due to withdrawal from the study treatment method or study:

Every individual participates in a safety follow-up visit 28 days after receiving the final dose of study treatment. Persons who discontinue investigating the product early may be requested to return to the hospital for an early termination visit and may be able to enter follow-up assessments. Early investigations The main reason for product discontinuance should be documented in the appropriate electronic case report format (eCRF).

Treatment of objects

Treatment to be administered: Compound 1 is administered in a 100 mg capsule to a dose of 200 to 300 mg (the planned dose level) three times a day (resulting in a daily dosage of 600 mg to 900 mg). Additional dose levels determined by the study SMC can be investigated. The study medication was administered to subjects entering the first visit (0 day), the third visit (28 days), the fourth visit (42 days), the fifth visit (56 days) and the sixth visit (84 days) Packed in # 2 capsules (100 mg) for each dispense at subsequent visits of the catheter, and then closed with a polyethylene cap liner installed in the cap. Research drugs are labeled according to national requirements. Several doses of anthraquinolone are planned for the first dose-increasing portion of the experiment. Dose increases follow the 3 + 3 design based on DLT occurrence. The starting dose of anthraquinolone is 200 mg three times a day, which is the dose currently being studied in an ongoing Phase II single agent trial in NSCLC. If there is no DLT, or less than 1 of 6 individuals have DLT, the dose is increased to 300 mg three times a day. One dose (MTD or MFD) is studied in the population expansion portion of the experiment.

An individual will receive anthraquinolone when judged by an investigator unless the individual continues to benefit clinically and has no cessation criteria. Anthraquinol should be taken every eight hours after about 15 minutes of eating a meal or light snack, and should not be consumed within ± 1 hour of drinking an ethanol-containing beverage, such as an alcoholic drink. Individuals who have forgotten or are unable to administer the dose at the scheduled time should be instructed to administer the dose as soon as possible. When they do not remember or can not administer doses prior to the next dosing schedule, they should take the planned doses and do not supplement the missed doses. The date and time of administration of each study drug should be recorded in the individual's journal.

The recommended dose of nab-paclitaxel in combination with gemcitabine is 125 mg / m ² via IV infusion over a period of 30 minutes on days 1, 8 and 15 of each 28-day cycle. The current recommended dose (RD) of gemcitabine is 1000 mg / m &lt; ² &gt; through IV infusion over 30 minutes immediately after the end of the Nab-paclitaxel administration on days 1, 8, and 15 of each 28- . Treatment with nab-paclitaxel + gemcitabine lasts until unacceptable toxicity or disease progression. In accordance with clinical practice and local rules, in the case of patient toxicity, the 15 day treatment dose may be withdrawn. In this case, the onset of the subsequent cycle begins early on the 22nd day of the first cycle administration (except cycle 1, DLT period). The tumor assessment plan remains unchanged (that is, the assessment is performed consistently every 8 weeks).

Selection of dosage for each individual and time of administration: In the dose-increasing portion of the study, individuals are registered and continuously treated according to a 3 + 3 dose-increasing design. In the collective enlargement, individuals are registered and treated simultaneously.

Efficacy and Safety Variables

Efficacy Assessment: Use the Resist 1.1 criteria to assess individual response to treatment by determining ORR, DoR, DCR, PFS, PFS 3 months, and PFS 6 months. Non-measurable, non-measurable, target and non-target lesion recurrence 1.1 guidelines and objective tumor response criteria (complete) based on the modified response criteria of the Solid Arm Cancer Instruction Manual Version 1.1 (supplemented with supplementary explanations and adapted from the original publication) Reactive [CR], partial response [PR], stable disease [SD] or progressive disease. OS, OS 6 months, and OS 12 months.

Baseline assessments should be performed within 28 days prior to initiation of study therapy and ideally should be as close as possible to initiation of study treatment. Every 8 weeks for the first 12 months, and every 12 weeks thereafter, the efficacy of all individuals is assessed by objective tumor assessment until objective disease progress is identified. If an unscheduled evaluation is performed and the entity does not proceed, an attempt should be made to perform a follow-up evaluation at the scheduled visit.

After a progressive disease is first demonstrated, a confirmatory scan is required. A confirmatory scan should preferably be performed at the next scheduled visit, and should be performed no later than four weeks after the initial evaluation of the progressive disease, without clinically significant deterioration. Continuous treatment is performed between the initial assessment of progress and confirmation of progress. If an individual discontinues treatment (and / or receives subsequent chemotherapy) before proceeding, the individual must continue to follow up until objective disease progress is identified.

An objective tumor response (CR or PR) should preferably be identified at the next scheduled visit and at least four weeks after the visit when the response is first observed.

After confirming the progress, the subject must continue to follow up every two months (8 weeks) to survive. In addition, all individuals should be contacted within one week after data cutoff to ascertain survival status.

Safety assessment: Safety is monitored throughout the study for all subjects. Analysis of the safety data is carried out using a Safety Analysis Set.

Adverse reactions:

Definition: The term "adverse event" used by the sponsor is synonymous with the term "adverse experience" as used by the FDA.

AE is any undesired, unwanted, unplanned clinical response in the form of signs, symptoms, illnesses, or laboratory or physiological observations that occur in a person participating in clinical studies using sponsored test products, regardless of causality. This includes:

● any clinically significant deterioration of existing disease

● Note: Expression of new pathogens related to clinical treatment during treatment at sites outside the initial infection site is considered as AE.

● Any recurrence of existing disease

• AEs resulting from overdose of accidental or intentional sponsored study drugs (ie, higher doses than those prescribed by health care professionals for clinical reasons)

• AEs resulting from abuse of sponsored study drugs (ie, use of non-clinical reasons)

• AE related to discontinuing use of sponsored study drugs

Note: The procedure is not AE, but the reason for the procedure may be AE.

Existing diseases are clinical diseases (including those being treated) that are diagnosed before the individual signs the ICF and are documented as part of the individual's medical history.

Using the question as to whether the disease was present prior to the onset of the active phase of the study and whether the disease has increased in severity and / or frequency, it is determined whether the response is a TEAE. AE is not present when (1) the active phase of the study is initiated, is not a chronic disease that is part of the individual's medical history, (2) exists at the initiation of the active phase of the study or exists as part of the individual's medical history If the severity or frequency is increased during the active period, AE is considered to be caused by treatment. The active phase of the study begins when the study drug is first administered. The active phase of the study is terminated upon subsequent follow-up visits.

Report of adverse events: At each visit, the investigator or the representative determines whether any AE has occurred. Individuals are questioned in the usual way and do not suggest specific symptoms. If any AEs are generated, they are recorded in the AE portion of the eCRF and in the medical record of the individual. If known, the diagnosis should be recorded before listing the individual signs and symptoms.

Adverse event reports begin at the time of the known consensus and end 30 days after the final administration of IMP.

Assessment of Severity: To assess AE severity, use the AE Severity Scoring Rating of NCI CTCAE (version 4.03). For AEs not specifically listed in the NCI CTCAE, use the following definition:

● Class 1 : Weak; Asymptomatic or mild symptoms; Clinical or diagnostic observations only; Or intervention not directed.

● Grade 2 : Medium; Minimal, local, or non-invasive intervention is indicated; Or age-appropriate means of daily living.

● Class 3 : Serious or medically significant, but not immediately fatal; Hospitalization or hospitalization is ordered; Disability; Or limits the ability of self care in daily life.

● Class 4 : Fatal consequences or emergency intervention indicated.

● Class 5 : AE related deaths.

Relationship to research therapy: The investigator determines the relationship of the AE to the study drug using a four-category system according to the following guidelines:

• Unrelated : A clinical response that has a temporal relationship incompatible with drug administration and can be explained by a baseline disease or other drug or compound, or is unrelated to the study drug.

• Low likelihood : A clinical response that creates a causal relationship that can not be time-correlated to drug administration, but can be reasonably explained by underlying disease or other drugs or compounds.

• Possible : clinical response that has a reasonable time relationship to the study drug, but can also be explained by coexisting disease or other drug or compound.

• Certain : Clinical responses that have a reasonable time relationship to study drug administration and can not be explained by coexisting disease or other drugs or compounds.

Follow-up of adverse events: When the investigator decides on the causal relationship, it is important to consider the time relationship of the onset of response to the underlying disease, the coexisting drug and the study drug, and the information on the outcome of the trial. Six months after the final administration of the study drug or discontinuation of follow-up (whichever occurs earlier), follow-up with the results of each AE until the disease is resolved or stabilized, and alternative treatment for pancreatic cancer is initiated It is the responsibility of the investigator to do this. If there is a serious or research drug-related toxicity, trace the individual until it is resolved or stabilized. Safety follow-up data can be collected by telephone contact every three months after the study visit is terminated.

Serious Adverse Reactions: A Serious Adverse Reaction (SAE) is any AE that occurs at any dose that meets one or more of the following criteria:

● Going to death

● Fatal (see below)

● Requires hospitalization of an individual or extension of existing hospitalization (see below)

● Continuous or significant inability or incapacity (see below)

● Causes deformities or congenital defects

● Causes significant medical reactions (see below)

It is also possible, on the basis of appropriate medical judgment, not to lead to death, not to be fatal, or to be hospitalized if it may be necessary to have medical or surgical intervention to prevent an individual from being listed and to prevent one of the listed results An important medical response that is not needed can be considered an SAE. Examples of such reactions include allergic bronchospasm requiring an intensive care roll at the emergency room or at home, blood diseases or convulsions that do not require hospitalization, or the occurrence of drug dependence or drug abuse.

Fatal AEs are arbitrary AEs in which an individual is at risk of immediate death therefrom when a reaction occurs. A fatal reaction does not include a reaction that causes death when it occurs in a more severe form, but does not pose an immediate risk of death when it actually occurs. For example, drug-induced hepatitis, which resolves without evidence of liver failure, is not considered fatal, even if the more serious nature of drug-induced hepatitis may be fatal. It is considered hospitalization only when approved for one night.

Admission or extension of hospitalization is a criterion that AE considers to be serious. If there is no AE, the participating investigator may not report admission or admission extension.

Also, planned hospitalization prior to commencement of study due to undiagnosed existing disease (eg, selective hospitalization for total knee replacement due to pre-existing osteoarthritis of the knee not worsened during the study) does not constitute an SAE.

Disability is defined as a substantial decline in the ability of a person to perform a function of daily living. If there is no doubt that the case constitutes AE or SAE based on the information available, this case should be treated as SAE.

Alternatively, medical and scientific judgments should be made in deciding whether the critical medical response is immediate, fatal, or whether the case is serious in situations where it does not result in death, hospitalization, disability or incompetence. These include reactions that may make the individual dangerous or may require medical intervention to prevent one or more consequences listed in the serious definition.

Pharmacokinetics  analysis

Non-compartment PK analysis is performed on individual plasma concentration data. PK analysis is performed using commercially available software such as Pheonix ™ WinNonlin® version 6.4 or higher (Certara USA, Inc.). The maximum plasma concentration (C _max ), the lowest (pre-dose) concentration (C _trough ), and the time of C _max (T _max ) are taken directly from the observed data. Using the linear trapezoidal formula, calculate the area under the plasma concentration-time curve (AUC _τ ). PK parameters for each individual are listed and summarized by treatment group using descriptive statistics (sample size [N], arithmetic mean, standard deviation, coefficient of variation [CV%], median, min, Geometric mean). Individual concentration data are listed and summarized by treatment group using descriptive statistics (N, arithmetic mean, standard deviation, median, min, max, geometric mean, and CV%). Average and individual plasma concentration-time profiles are provided graphically on a linear scale and a semi-log scale.

The actual dose administration time and sample collection time are used for analysis recorded in the case report form. Plasma concentrations below the lower limit of quantification are set to zero in this analysis. Pharmacokinetic parameter estimates are three or four significant numbers for presentation. And does not attempt to estimate missing data. Other parameters and data handling procedures may be added where appropriate. Only those individuals with active anthraquinolone provided and having a valuable concentration-time profile are included in the analysis. All statistical analyzes use untested parameter estimates.

A population-based PK (PopPK) model can be developed and analyzed using a mixed-effect method. Combine administration, sampling, statistics and laboratory data from each individual into a database suitable for population analysis. We develop a nonlinear mixed-effect model using NONMEM (Globomax, Ellicott City, Maryland). The various structural models are evaluated as indicated by the data. Covariate effects (age, sex, race, body size, smoking history, etc.) are assessed and incorporated into the model using a univariate or multivariate approach. Once an optimal model is established, it is validated using standard methods such as visual prediction checks and bootstrap analysis.

Exposure-exposure / concentration relationships are evaluated based on the post-analysis estimates (ie, model predicted C _max , C _trough and AUC _τ ) of anthraquinone exposure from the PopPK model or concentration observation.

Statistical method

General considerations: For research reporting purposes, consider the following aspects:

● Technical statistics and graphical presentations are the main analytical tools.

● Report individual batches for each dose level using all individuals in the two phases of the study and provide individual profiles only for the dose increments of the study.

• For all analyzes, the results and graphical representations of individual entity data are provided only at the dose level. The dose level assigned to the subject at the time of initial treatment is displayed on all outputs.

Using descriptive statistics and graphical representations, summarize the data for each dose level. Using the following summary statistics, unless otherwise specified, summarize the experimental data per dosage level based on the characteristics:

● Continuous variables: Nondeterministic (N), mean, standard deviation, median, minimum and maximum; Where appropriate 95% CI is provided.

● Category variables: frequency and percentage.

Details of the statistical analysis are provided in the statistical analysis plan.

Efficacy analysis

Main results measurement:

Dose increasing step : The number and percentage of individuals experiencing DTL at each dose level during cycle 1 are evaluated on a DLT assay set.

Population Expansion Phase : Best overall response in each category (CR, PR, SD, and progressive disease) with a 95% CI according to rescue 1.1; Objective response rate (CR + PR); A technical summary of the subjects with disease inhibition rates (CR + PR + SD [SD for 16 weeks or more]) is performed on the efficacy analysis set.

The baseline (resis) version 1.1 in solid tumors was performed for intermediate OS time and the PFS time with 95% CI was assessed using the Kaplan-Meier method on the entire assay set. The OS is defined as the time from the initial administration of the study drug to the death of the individual. PFS is defined as the time from the initial administration of the study drug to the onset of disease progression or to the mortality (whichever comes first).

Analysis of secondary outcome measures: Describe secondary endpoint data by dose level, subject and visit (if applicable). The secondary endpoints are characterized by dosage levels in the relevant assay set.

Safety and tolerability analysis : Number and proportion of individuals experiencing TEAE; Drug exposure; Clinically significant changes in laboratory parameters, vital signs, physical examination, body weight, ECOG performance status, ECG and / or pulse oximetry, as judged to be related to the experimental medication; And the number of deaths and reasons.

Safety assessments include the occurrence of AE (or TEAE), laboratory test results, vital signs, ECG results, and physical examination. All summaries of safety data are based on a set of safety analyzes. No formal statistical analysis of safety data is performed.

A summary table is provided for all adverse events (AEs) by dose level, and medical pre-SOC and preferred terms for regulatory work where applicable. The occurrence and type of AEs are analyzed.

AE is thought to be "caused by treatment " when it occurs after the initial study treatment or before the first study treatment and then deteriorates.

The TEAE summary table contains the number of objects. Thus, when an entity experiences more than one episode of a particular AE, the entity is counted only once for that reaction. If an entity has more than one AE coded in the same preferred term, the entity is counted only once for that preferred term. Similarly, if an entity has more than one AE in the SOC, the entity is counted only once in that SOC.

The cause of death as well as all deaths and deaths within 30 days of the last dose of study treatment are tabulated by dose level. Drug exposure is summarized by dose level.

The clinically significant laboratory test parameters were calculated from descriptive statistics (number of individuals, mean, standard deviation, minimum value, maximum value, and mean change from baseline, standard deviation for mean and standard error, , The maximum value, and the number and percentage of individuals within a defined category). Cross-tabulation of variance tables (ie baseline versus less than the lower limit of the normal range at the planned visit, within the normal range, and above the upper limit of the normal range) is provided by laboratory tests (where applicable). Laboratory tests with category results that can not be analyzed by baseline or by change table analysis are not included in these summaries, but are listed. Data obtained from laboratory tests not required by the experimental method are listed, but not summarized.

Descriptive statistics of life signs, weight, and ECG results at each visit are provided by the treatment group. Physical examinations are listed for each individual. The overall response rate and DCR are predicted, along with the 95% CI calculated using the Clopper and Pearson methods by dose level.

drug Dynamic  analysis

Pharmacokinetic sampling: PK samples are taken at day 0 and day 28 in all subjects enrolled at stage 1 at this point:

Day 0: 30 minutes before the first dose, and 0.5, 1, 2, 3, 4, 6, and 8 hours after the first dose (about 5 mL per sample, total 60 mL).

Day 28: immediately before the first dose on day 28, and after 0.5, 1, 2, 3, 4, 6, and 8 hours after the initial dose (about 5 mL per sample, total 60 mL).

Perform lean PK sampling at day 28, day 42, and day 56 on all subjects enrolled in step 2. Two or more samples are collected in each case, one of which is the lowest concentration (30 minutes before the first dose on days 28, 42 and 56, and about 8 hours after the last dose on the previous day). The time is adjusted so that at least one sample per subject will match the peak concentration (after 3 hours of initial administration). The remainder can be taken at any time during the dosing interval.

Each blood sample is analyzed for anthraquinolone plasma concentration to determine the PK parameters after administration of anthraquinolone using a fully-approved bioanalytical method (a copy of the analysis methodology report and a bioanalytical report included in the clinical study report do).

drug Dynamic  Sample collection procedure

Blood sample: A venous blood sample is obtained from a 5 mL sodium heparin Vacutainer® tube at a predetermined sample time. Immediately after collection, the tube is gently inverted eight to ten times to mix the blood sample with the anti-caking agent. All samples are processed and placed in the freezer within 1 hour of collection. Plasma fractions are separated by placing the collection tube at 3000 rpm for 10 minutes in a refrigerated centrifuge (4 ° C). The plasma fraction is recovered by pipette and divided into two polypropylene freezing tubes (each tube receives approximately the same aliquot). All sample collections and frozen tubes are clearly labeled by object number, study period and collection time. The label is fixed to the freezing tube in such a way as to prevent the label from falling off after freezing. All plasma samples are placed in a -70 ° C freezer within 1 hour of collection.

Analytical Method: Determine the concentration of the study drug from the plasma sample using approved analytical methods. Details of method approval and sample analysis are included in the final clinical study report.

Preliminary results:

Preliminary results from patients receiving a 600 mg daily dose show that patient tumor size (mm) decreased after the third cycle (8 weeks of treatment) was terminated. The tumor size of one patient was reduced by 7.6% from 91mm to 84mm, and the tumor size of the other patients was reduced by 22% from 50mm to 39mm. The preliminary tumor size reduction may be further characterized by the addition of an excellent and unexpected advantage of the compositions of the present invention including pancreatic cancer, current clinical chemotherapeutic drugs such as paclitaxel and gemcitabine, and Exemplary Compound 1, Backed.

While preferred embodiments of the invention have been illustrated and described herein, those skilled in the art will readily appreciate that such embodiments are provided by way of example only. Those skilled in the art will now make numerous modifications, changes, and substitutions without departing from the invention. It should be understood that various alternatives to the embodiments of the invention described herein can be utilized in practicing the invention. The following claims are intended to define the scope of the invention, and the methods and structures within the scope of these claims, and equivalents thereof, are intended to be encompassed by the claims.

Claims (20)

The

Wherein R ₁ , R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, methyl or (C ₁ -C ₈ alkyl) CH _{2) m} -CH _3, and; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C aryl optionally substituted with one or more substituents selected from ₈ haloalkyl; n is the structure of 1 to 12 Im) Or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof, for the manufacture of a medicament for treating pancreatic cancer in a subject. The method according to claim 1,
Wherein said at least one anticancer agent is selected from the group consisting of gemcitabine, paclitaxel, dirubicin / cytarabine, ethocidephosphate, gleevec, temozolomide, bortezomib, letrozole, cetuximab, bevacizumab, nab-paclitaxel, docetaxel, Pemetrexed / Carboplatin, Retrosol / Cyclophosphamide, Temsirolimus, Bevacizumab / Temsirolimus, Filimovab, RAD001, Pazopanie, Pemetrexed / Porphyromycin, cisplatin, cytarabine / daunorubicin, tallirolimus, allotinib / temsilolorimus, paclitaxel, paclitaxel, Selected from the group consisting of capecitabine, tamoxifen, bortezomib, trastuzumab, docetaxel / capecitabine, trastuzumab / tipiparnib, tipiparnib / gemcitabine, / RTI &gt; The method according to claim 1,
Wherein the at least one anticancer agent is gemcitabine, paclitaxel, or a combination thereof. The method according to claim 1,
Wherein the composition further comprises an immunotherapeutic agent. 5. The method according to any one of claims 1 to 4,
Wherein X and / or Y is oxygen. 5. The method according to any one of claims 1 to 4,
Wherein R is _{hydrogen, C (= O) C 3} H 8, C (= O) C 2 H 5 or C (= O) CH ₃ in the composition. 5. The method according to any one of claims 1 to 4,
Wherein each of R ₁ , R ₂ and R ₃ is independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl or octyl. 5. The method according to any one of claims 1 to 4,
The compound

/ RTI &gt; A pharmaceutical composition according to claim 1 for use in treating pancreatic cancer in an individual. For use in the treatment of patients with pancreatic cancer who are resistant to, not treated with, or nonresponsive to, gemcitabine, paclitaxel, or a combination thereof.

Wherein R ₁ , R ₂ and R ₃ are each independently selected from the group consisting of hydrogen, methyl or (C ₁ -C ₈ alkyl) CH _{2) m} -CH _3, and; R ₄ is H, C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, or C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl, and C ₁ -C aryl optionally substituted with one or more substituents selected from ₈ haloalkyl; n is the structure of 1 to 12 Im) Or a pharmaceutically acceptable salt, metabolite, solvate, or prodrug thereof. 11. The method according to claim 9 or 10,
Wherein said compound, or a pharmaceutically acceptable salt, metabolite, solvate or prodrug thereof, is administered orally, parenterally, intravenously or by injection. 11. The method according to claim 9 or 10,
Wherein the composition further comprises an immunotherapeutic agent. 11. The method according to claim 9 or 10,
Wherein X and / or Y is oxygen. 11. The method according to claim 9 or 10,
Wherein R is _{hydrogen, C (= O) C 3} H 8, C (= O) C 2 H 5 or C (= O) CH ₃ in the composition. 11. The method according to claim 9 or 10,
Wherein each of R ₁ , R ₂ and R ₃ is independently hydrogen, methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl or octyl. 16. The method of claim 15,
Wherein R &lt; _{1 &gt;} is hydrogen or methyl. 16. The method of claim 15,
Wherein R &lt; _{2 &gt;} is hydrogen or methyl. 11. The method according to claim 9 or 10,
Wherein R ₄ is H or _one or more selected from C ₁ -C ₈ alkyl, C ₂ -C ₈ alkenyl, C ₂ -C ₈ alkynyl, C ₃ -C ₈ cycloalkyl or C ₁ -C ₈ haloalkyl, C ₁ -C ₈ alkyl optionally substituted with a substituent of the composition. 19. The method of claim 18,
Wherein R ₄ is _{CH 2 CH = C (CH 3} ) 2 composition. 20. The method of claim 19,
The compound

/ RTI &gt;

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