KR20230111071A - Adjuvant for Radioimmunotherapy of Cancer Comprising Lenvatinib - Google Patents

Abstract

The present invention relates to a radioimmunotherapy adjuvant for cancer containing lenvatinib as an active ingredient, and when administered in combination with the radioimmunotherapy agent and lenvatinib, the effect of radioimmunotherapy is remarkably improved. Therefore, the present invention can be usefully used for cancer treatment because radioimmunotherapy minimizes the effect on normal cells while providing effective anticancer treatment.

Description

Adjuvant for Radioimmunotherapy of Cancer Comprising Lenvatinib

The present invention relates to an adjuvant for cancer radioimmunotherapy and a composition for enhancing radioimmunotherapy containing lenvatinib as an active ingredient.

BACKGROUND OF THE INVENTION Cancer is a disease that causes malignant tumors by unlimited proliferation of cells in biological tissues, and eventually invades surrounding tissues or metastasizes to other organs, leading to death of the living body. In addition to hereditary causes, physical stimulation, chemical stimulation, and viral infection are the causes, and it is difficult to cure.

Cancer treatment methods include radiation therapy, chemotherapy using anticancer agents, and radioimmunotherapy in which tumor-specific antibodies and therapeutic radioactive isotopes are combined. Treatment using radiation has a high frequency of side effects, and there is a problem that its use is limited. In particular, when it is difficult to treat through surgery, the treatment is performed using radiation. In order to minimize damage to normal organs and increase the treatment effect, radiation should be adjusted according to the stage and location of cancer. Although there is a difference in sensitivity between individuals or tissues during radiation therapy for cancer cells, if the sensitivity to radiation can be increased, the same therapeutic effect can be obtained by irradiating a patient with a smaller amount of radiation.

Radioimmunotherapy (RIT) is a treatment that can maximize the therapeutic effect by labeling a tumor epitope-specific monoclonal antibody with a therapeutic radioisotope (I-131, Y-90, Lu-177, etc.) that emits beta rays, and using a combination of anti-cancer effects of monoclonal antibodies and cytotoxic radiation. Radioimmunotherapy can expect a synergistic effect of cancer treatment by targeted therapy of tumor antibodies and cancer treatment effect by radiation. Accordingly, studies on radioimmunotherapy are being actively conducted, but there are also cases in which the response of radioimmunotherapy is low depending on the type or progress of some patients or cancer.

Lenvatinib is a targeted anticancer drug that inhibits the growth of cancer cells by binding to signal transduction receptors such as vascular endothelial growth factor receptors (VEGFR) 1, 2, 3 and platelet-derived growth factor receptor alpha to block signals necessary for survival and division of cancer cells. Lenvatinib (trade name: Lenvima) is used in the treatment of certain types of thyroid cancer and hepatocellular carcinoma, but it is known that the anticancer effect of lenvatinib alone is limited because no anticancer effect is seen in more than 60% of patients. International patent application PCT/JP2014/067723 (WO2014-208774) discloses a composition for anticancer treatment using lenvatinib and eribulin in combination. However, no literature has been reported yet showing that lenvatinib can improve the therapeutic effect in radioimmunotherapy using radioimmunotherapy.

PCT/JP2014/067723 (WO2014-208774)

As a result of research efforts to develop a combination drug that can increase the cancer therapeutic effect of radioimmunotherapeutic agents, the inventors of the present invention completed the present invention by experimentally confirming that the therapeutic effect of radioimmunotherapeutic agents increases synergistically when lenvatinib is used in combination with radioimmunotherapeutic agents.

The present invention provides an adjuvant for cancer radioimmunotherapy comprising lenvatinib as an active ingredient.

In addition, a composition for enhancing cancer radioimmunotherapy comprising lenvatinib as an active ingredient is provided.

In addition, the present invention provides a pharmaceutical composition for treating or preventing cancer comprising lenvatinib and a radioimmunotherapy as active ingredients.

One aspect of the present invention provides an adjuvant for cancer radioimmunotherapy comprising lenvatinib as an active ingredient.

The lenvatinib is a compound having a structure represented by Chemical Formula 1 below, and can be purchased commercially and used or prepared by a chemical synthesis method known in the art.

[Formula 1]

The lenvatinib may be used in the form of a salt, particularly a mesylate. Lenvatinib is a targeted anticancer agent that inhibits the growth of cancer cells by inhibiting receptor tyrosine kinases such as vascular endothelial growth factor receptor (VEGFR), and is used in the treatment of thyroid cancer and hepatocellular carcinoma.

In one embodiment, the cancer radioimmunotherapy adjuvant containing lenvatinib as an active ingredient of the present invention may be administered in combination with a radioimmunotherapy agent.

The combined administration of the adjuvant may be concurrently administered with the radioimmunotherapy agent, separately or sequentially.

The cancer radioimmunotherapy adjuvant containing lenvatinib as an active ingredient can be administered in combination with a radioimmunotherapy agent, thereby synergistically enhancing the anticancer effect of the radioimmunotherapy agent.

The adjuvant of the present invention may be one that exhibits a synergistic therapeutic effect with the radioimmunotherapy agent or enhances the anticancer effect of the radioimmunotherapy agent for cancer in anticancer therapy using the radioimmunotherapy agent. Enhancement of the anticancer effect of the radioimmunotherapy agent for cancer may be to improve the therapeutic sensitivity or radiosensitivity of the radioimmunotherapy agent. Therapeutic sensitivity or sensitivity of a radioimmunotherapy agent means the degree of therapeutic response to radiation therapy, and high sensitivity or sensitivity is described when a high therapeutic effect is exhibited for a small amount of radiation. In the present invention, when an adjuvant is used in combination with a radioimmunotherapeutic agent, even if a smaller dose of the radioimmunotherapeutic agent is treated than when the adjuvant is not used in combination, it may mean that an anticancer effect equivalent to the case of using a larger amount of the radioimmunotherapeutic agent appears. Alternatively, when the adjuvant is used in combination with a radioimmunotherapeutic agent, it may mean that a superior anticancer effect is exhibited as compared to the use of an equivalent dose of the radioimmunotherapeutic agent alone.

In the present invention, radioimmunotherapy (RIT) is a type of radiation therapy in which a radioactive material is combined with an immunotherapeutic agent and injected into the body, and is an immunotherapeutic agent that can specifically bind to a target antigen located on the surface of cancer cells. For example, it is a targeted radiotherapy method in which a radioactive isotope is bound to a monoclonal antibody. Radioimmunotherapy has the advantage of being able to specifically treat cancer cells with a target antigen, thereby minimizing the effect on normal cells and increasing the anticancer effect.

In the present invention, the radioimmunotherapy agent may be a drug used for radioimmunotherapy, specifically an antibody paired with a radioactive material called a radiotracer, and more specifically an antibody labeled with a radioactive isotope.

The antibody is a polypeptide comprising a framework region from an immunoglobulin gene or fragment thereof that specifically binds and recognizes an antigen, and includes, for example, monoclonal antibodies, polyclonal antibodies, chimeric antibodies, humanized antibodies, and human antibodies. The monoclonal antibody refers to an antibody derived from a single B cell clone, and the polyclonal antibody refers to an antibody produced and secreted from different B cell clones. The chimeric antibody refers to an antibody molecule artificially formed by processing two types of antibodies against different antigens so that one molecule binds to both antigens. In addition, the humanized antibody is an antibody that retains the reactivity of a non-human antibody while having low immunogenicity to humans. Such humanized antibodies can be obtained, for example, by retaining non-human CDR regions and substituting the remaining portions of the antibody with their human counterparts. The antibody includes not only a complete antibody form, but also an antigen-binding fragment (antibody fragment) of an antibody molecule. A complete antibody is a structure having two full-length light chains and two full-length heavy chains, each light chain linked to the heavy chain by disulfide bonds. The heavy chain constant region has gamma (γ), mu (μ), alpha (α), delta (δ) and epsilon (ε) types, and subclasses include gamma 1 (γ1), gamma 2 (γ2), gamma 3 (γ3), gamma 4 (γ4), alpha 1 (α1) and alpha 2 (α2). The constant region of the light chain has kappa (Κ) and lambda (λ) types. An antigen-binding fragment of an antibody or antibody fragment refers to a fragment having an antigen-binding function, and includes Fab, F(ab'), F(ab')2, Fv, and the like.

In one embodiment, the radioimmunotherapy agent may be a radioisotope-labeled anti-HER2 antibody. The anti-HER2 antibody may be Trastuzumab. Trastuzumab (trade name: Herceptin) binds specifically to HER2, which is involved in tumor growth, and can exert a pharmacological action on metastatic and early breast cancer in which the human epidermal growth factor receptor 2 (HER2) gene is overexpressed.

The radioactive isotope refers to an isotope that decays into a stable element by emitting radiation such as alpha particles, neutrons, beta rays, gamma rays, and X-rays. Among them, isotopes that emit alpha particles, beta rays, and gamma rays have low penetrating power and excellent cell ionization ability, so they can be used for medical purposes. The radioisotope may be an alpha particle emitting radioisotope, a beta ray emitting radioisotope or a gamma ray emitting radioisotope, for example, Cu-67, Y-90, Pb-103, Sn-117m, Ho-166, I-131, Lu-177, Re-188, At-211, Bi-213, Ra-223, Ac-225, Th-227, or In-1 11, specifically I-131 ( ¹³¹ I).

In one embodiment, the radioisotope-labeled anti-HER2 antibody may be ¹³¹ I-trastuzumab.

In the present invention, cancer, which is a disease to be treated in the present invention, generally refers to a physiological state of a mammal having abnormal cell growth characteristics, and abnormal overproliferation due to a problem in the regulation function of normal division, differentiation, and death of cells. Infiltrate into surrounding tissues and organs to form lumps and destroy or transform existing structures. The cancer may be any one cancer selected from the group consisting of blood cancer, breast cancer, head or cervical cancer, uterine cancer, cervical cancer, ovarian cancer, lung cancer, bladder cancer, esophageal cancer, neoplastic tumor, neuroblastoma, testicular cancer, leukemia, stomach cancer, liver cancer, colon cancer, skin cancer, brain cancer, laryngeal cancer, prostate cancer, thyroid cancer, kidney cancer, and rectal cancer. The cancer may be HER2 (human epidermal growth factor receptor 2) positive cancer, HER-positive breast cancer or cancer metastasized from the breast cancer, HER2-positive gastric cancer, lung cancer, hematological cancer, lymphoma, follicular lymphoma, diffuse large B-cell lymphoma, mantle cell lymphoma, B-CLL/SLL, immunocytoma/Wal denstrom's), MALT-type/monocytoid B cell lymphoma, Burkitt's lymphoma, pediatric lymphoma, and anaplastic large cell lymphoma, but is not limited thereto.

The 'anti-cancer effect' means prevention or treatment of the cancer. The prevention refers to any action that suppresses or delays the occurrence or progression of cancer, and the treatment means not only death of cancer cells, but also all effects in which cancer cells do not get worse, such as reduction in growth, reduction in proliferation, reduction in recurrence, alleviation of one or more symptoms associated with cancer to some extent, formation of immune system anticancer activity and immune memory formation in cancer cells, or reduction in invasion of cancer cells, but are not limited thereto.

The adjuvant of the present invention may be provided in the form of a pharmaceutical composition. In this case, the pharmaceutical composition may further include an active ingredient exhibiting anticancer activity in addition to lenvatinib. The pharmaceutical composition may further include a pharmaceutically acceptable carrier or additive in addition to the active ingredient.

The meaning of 'pharmaceutically acceptable' means that it does not inhibit the activity of the active ingredient and does not have toxicity more than is adaptable to the subject of application (prescription). The carrier is defined as a compound that facilitates the addition of the compound into cells or tissues.

The pharmaceutical composition may be administered by mixing with any convenient carrier, etc., and such dosage form may be a single or repeated dosage form. The composition may be a solid formulation or a liquid formulation. Solid preparations include, but are not limited to, powders, granules, tablets, capsules, and suppositories. Solid formulations may include carriers, flavoring agents, binders, preservatives, disintegrants, lubricants, fillers, and the like, but are not limited thereto. Liquid preparations include solutions, suspensions, and emulsions such as water and propylene glycol solutions, but are not limited thereto, and may be prepared by adding appropriate colorants, flavoring agents, stabilizers, viscosifiers, and the like. For example, a powder may be prepared by simply mixing an appropriate pharmaceutically acceptable carrier such as lactose, starch, or microcrystalline cellulose in addition to genipin, which is an active ingredient of the present invention. Granules can be prepared by mixing the active ingredient of the present invention with a suitable pharmaceutically acceptable carrier and a suitable pharmaceutically acceptable binder such as polyvinylpyrrolidone or hydroxypropylcellulose, and then using a wet granulation method using a solvent such as water, ethanol, or isopropanol or a dry granulation method using compression force. In addition, tablets may be prepared by mixing the granules with a suitable pharmaceutically acceptable lubricant such as magnesium stearate and then tableting them using a tableting machine. When formulating the pharmaceutical composition as an injection, it may be prepared according to a conventional method for preparing an injection known in the art. When formulated as an injection, it may be dispersed in a sterile medium so that it can be used as it is when administered to a patient, or may be administered after dispersing in an appropriate concentration by adding distilled water for injection.

The pharmaceutical composition may be administered as an injection (e.g., intravenous injection, intramuscular injection, intraperitoneal injection, infusion, subcutaneous injection, implant), inhalant, oral agent, nasal agent, vaginal agent, rectal agent, sublingual agent, transdermal agent, topical agent, etc., depending on the disease to be treated and the condition of the subject, but is not limited thereto. Depending on the route of administration, it can be formulated into an appropriate dosage unit formulation containing a pharmaceutically acceptable carrier, excipients, and vehicles that are commonly used and non-toxic. Suitable pharmaceutically acceptable carriers and agents are described in detail in Remington's Pharmaceutical Sciences ( ^19th ed., 1995).

The pharmaceutical composition may be administered at about 0.0001 mg/kg (body weight) to about 10 g/kg (body weight) daily, and at a daily dosage of about 0.001 mg/kg (body weight) to about 1 g/kg (body weight). The therapeutically effective amount or effective dose of the pharmaceutical composition may vary depending on the formulation method, administration method, administration time and/or route of administration, etc., and may vary depending on the type and degree of response to be achieved by administration of the composition, the type of subject to be administered, age, weight, general health condition, symptoms or severity of disease, sex, diet, excretion, drugs used simultaneously or at different times in the subject, and other components of the composition, etc., and similar factors well known in the pharmaceutical field. A person of ordinary skill in the art can readily determine and prescribe an effective dosage for the desired treatment. In addition, if necessary, for convenience, the total daily dose may be divided and administered several times during the day. The term “treatment-effective amount” refers to an amount sufficient to produce a desired effect, including treatment or improvement of cancer in a patient with cancer, side effects of anti-cancer treatment or resistance to anti-cancer drugs, improvement of a condition (e.g., one or more symptoms), delay of progression of a disease, and the like, in a patient with cancer.

Another aspect of the present invention provides a composition for enhancing cancer radioimmunotherapy comprising lenvatinib as an active ingredient.

The composition may be administered in combination with a radioimmunotherapy agent.

The radioimmunotherapy agent may be an antibody labeled with a radioactive isotope.

In addition, another aspect of the present invention provides a pharmaceutical composition for treating or preventing cancer comprising lenvatinib and a radioimmunotherapy as active ingredients.

The radioimmunotherapy agent may be an antibody labeled with a radioactive isotope.

In addition, another aspect of the present application provides a method for treating cancer by administering lenvatinib in combination with a radioimmunotherapy agent.

The radioimmunotherapy agent may be an antibody labeled with a radioactive isotope.

In the other aspect of the present invention, the composition for enhancing cancer radioimmunotherapy, the pharmaceutical composition for treating or preventing cancer, and the method for treating cancer, the related contents such as combination administration, Genipin, radioimmunotherapy, radioimmunotherapy, pharmaceutical composition, cancer, etc. are the same as those described in the adjuvant for cancer radioimmunotherapy, which is one aspect of the present invention described above, and in order to avoid excessive complexity of the specification, these are incorporated and will not be repeatedly described.

The adjuvant for radioimmunotherapy of cancer or the composition for promoting radioimmunotherapy of cancer containing lentatinib as an active ingredient of the present invention can enhance the anticancer effect of the radioimmunotherapy agent. According to the present invention, a desired therapeutic effect can be obtained even when a radioimmunotherapeutic agent is used at a reduced dose, thereby minimizing the radiation effect on normal cells and increasing the specific therapeutic effect on cancer cells.

However, the effects of the present application are not limited to the effects mentioned above, and other effects not mentioned will be clearly understood by those skilled in the art from the following description.

1 shows the incorporation yield of ¹³¹ I-trastuzumab produced by labeling trastuzumab with ¹³¹ I radioactive isotope.
Figure 2 shows the survival rate of NCI-N87 cells treated with ¹³¹ I-trastuzumab and lenvatinib at various doses.

Hereinafter, the present invention will be described in detail by examples. However, the following examples specifically illustrate the present application, and the content of the present invention is not limited by the following examples.

[Example 1]

culture of cells

Human gastric cancer cell lines were purchased from the Korea Cell Line Bank (KCLB) and used. 10% fetal bovine serum (FBS, Gibco) was added to RPMI-1640 medium (Corning, Inc., NY, USA) containing 1% antibiotic-antimycotic (100X, Gibco) and cultured. Cells were cultured at 37°C in a 5% CO ₂ atmosphere.

[Example 2]

¹³¹ Preparation of I-trastuzumab

Trastuzumab was purchased from Roche. For radioisotope labeling of the Trastuzumab, 1 mCi of ¹³¹ I was added to a Pierce Iodination tube (Thermo science, #28601) containing 1 mg of Trastuzumab and PBS (pH 7.4). After incubation at 25 ° C. for 15 minutes, the incorporation yield was calculated by thin layer chromatography using acetone (iTLC, Agilent Technologies). As a result, as shown in FIG. 1, the binding yield always exceeded 95%, and trastuzumab was successfully labeled with ¹³¹ I.

[Example 3]

¹³¹ Synergistic effect confirmed when using the combination of I-trastuzumab and lenvatinib

When ¹³¹ I-trastuzumab and lenvatinib were used in combination, an experiment was conducted to determine whether there was a synergistic effect of radioimmunotherapy.

Cell viability was measured using the EZ-Cytox cell viability, proliferation, and cytotoxicity assay kit (DoGenBio), and was performed according to the manufacturer's protocol. Specifically, NCI-N87 cells (HER2-positive) were treated with ¹³¹ I (10 μCi, 50 μCi, 100 μCi), ¹³¹ I-trastuzumab (10 μCi, 50 μCi, 100 μCi) alone, respectively, or 50 μM lenvatinib and ¹³¹ I-trastuzumab (10 μCi, 50 μCi, 1 00 μCi) were treated in combination. NCI-N87 cells (HER2 positive) were cultured for 48 hours at 37°C in a 5% CO ₂ atmosphere in a humidified incubator, then washed once with PBS, and then cultured for an additional 72 hours with the addition of 50 μM lenvatinib. Then, 10 μl of EZ-Cytox solution was added to each well and further incubated at 37°C for 3 hours in a CO ₂ incubator. Cell viability was quantified by absorbance at 450 nm using a SpectraMax i3 microplate reader (Molecular Devices, Sunnyvale, Calif.). All analyzes were repeated three times, and cell viability (%) was calculated as a percentage of the optical density (OD) ratio of the drug-treated and untreated samples, and the cell death effect of ¹³¹ I-trastuzumab and lenvatinib on NCI-N87 cell viability was evaluated.

As a result, as shown in Figure 2, the case of treatment with ¹³¹ I-trastuzumab and lenvatinib (47%, 39%, and 31%) increased cell death in NCI-N87 cells compared to the treatment with only ¹³¹ I (98%, 89%, 66%) or the treatment with only ¹³¹ I-trastuzumab (83%, 73%, and 59%). extinction could be ascertained. That is, when NCI-N87 (HER2-positive) cells were treated with ¹³¹ I-trastuzumab together with 50 μM lenvatinib, the cell viability was much lower than that of the same dose of ¹³¹ I or ¹³¹ I-trastuzumab alone. From the above results, it was confirmed that there is a synergistic anticancer effect when lenvatinib and ¹³¹ I-trastuzumab are used in combination.

Claims (12)

An adjuvant for cancer radioimmunotherapy comprising lenvatinib as an active ingredient.
The method of claim 1,
The adjuvant is a radioimmunotherapy adjuvant for cancer, which is administered in combination with a radioimmunotherapy agent.
The method of claim 2,
The adjuvant is a radioimmunotherapy adjuvant for cancer, which is administered concurrently, separately, or sequentially with the radioimmunotherapy agent.
The method of claim 2,
The radioimmunotherapy agent is a radioimmunotherapy adjuvant for cancer, which is a radioactive isotope-labeled antibody.
The method of claim 4,
The antibody is an anti-HER2 antibody, an adjuvant for cancer radioimmunotherapy.
The method of claim 4,
The radioactive isotope is Cu-67, Y-90, Pb-103, Sn-117m, Ho-166, I-131, Lu-177, Re-188, At-211, Bi-213, Ra-223, Ac-225, Th-227, and At least one selected from the group consisting of In-111, radioimmunotherapy adjuvant for cancer.
The method of claim 1,
The adjuvant for radioimmunotherapy of cancer, wherein the cancer is HER 2 (human epidermal growth factor receptor 2) positive cancer.
A composition for enhancing cancer radioimmunotherapy comprising lenvatinib as an active ingredient.
The method of claim 8,
The composition is a composition for promoting radioimmunotherapy of cancer, which is administered in combination with a radioimmunotherapy agent.
The method of claim 8,
The radioimmunotherapeutic agent is a radioisotope-labeled antibody, a composition for promoting radioimmunotherapy of cancer.
A pharmaceutical composition for the treatment or prevention of cancer comprising lenvatinib and a radioimmunotherapy as active ingredients.
The method of claim 11,
The radioimmunotherapy is an antibody labeled with a radioactive isotope, a pharmaceutical composition for the treatment or prevention of cancer.