JP2019156798A - Multiple myeloma therapeutic agent - Google Patents

Abstract

To provide a multiple myeloma therapeutic agent.SOLUTION: A multiple myeloma therapeutic agent has a synoviolin activity inhibitor as an active ingredient. Examples of the synoviolin activity inhibitor includes a naphthoquinone derivative having a specific substituent, a 1,3,5-triazine derivative having a specific substituent, or the like.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a therapeutic agent for multiple myeloma comprising an inhibitor of synoviolin activity as an active ingredient.

  In Table 2014/104224, Synoviolin activity inhibitors are described.

  Japanese Patent No. 4599501 describes a therapeutic agent for multiple myeloma containing a carboline derivative as an active ingredient.

  Japanese Patent No. 5665862 describes a therapeutic agent for bortezomib-resistant multiple myeloma.

No. 2014/104224 Japanese Patent No. 4599501 Japanese Patent No. 5665862

  An object of the present invention is to provide a therapeutic agent for multiple myeloma and a therapeutic agent for bortezomib-resistant multiple myeloma.

The present invention is basically based on an example in which an inhibitor of synoviolin activity is effective in the treatment of multiple myeloma.
That is, the present invention relates to a therapeutic agent for multiple myeloma, and includes a synoviolin activity inhibitor as an active ingredient.

（式（Ｉ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。
Ｘ^１及びＸ^２は，同一でも異なってもよく，酸素原子又は硫黄原子を表す。
Ａ^１−Ａ^２は，Ｃ−Ｃ（単結合），又はＣ＝Ｃ（二重結合）を表す。） An example of a synoviolin activity inhibitor is a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (I),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom.
A ¹ -A ² represents C—C (single bond) or C═C (double bond). )

（式（ＩＩ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。
Ｘ^１及びＸ^２は，同一でも異なってもよく，酸素原子又は硫黄原子を表す。） An example of a synoviolin activity inhibitor is a compound represented by the following formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (II),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom. )

（式（ＩＩＩ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。） An example of an activity inhibitor of synoviolin is a compound represented by the following formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (III),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom. )

Another example of a synoviolin activity inhibitor relates to the 1,3,5-triazine derivative represented by formula 1, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. An example of the 1,3,5-triazine derivative is a substance (LS-102) having the following chemical formula. LS-102 ^{is, (S) -N 2 - (} benzo [d] oxazol-6-yl) -N 4 - (1-cyclohexylethyl) -N 6 -ethyl-N 6 - (2- (ethylamino) ethyl) - 1,3,5-triazine-2,4,6-triamine ( (S) -N 2 - ( benzo [d] oxazol-6-yl) -N ⁴ - (1-cyclohexylethyl) -N ⁶ - ethyl - N ^6- (2- (ethylamino) ethyl) -1,3,5-triazine-2,4,6-triamine).

  The therapeutic agent for multiple myeloma of the present invention can be preferably used as a therapeutic agent for bortezomib-resistant multiple myeloma. For example, this therapeutic agent is preferably used as a therapeutic agent for multiple myeloma for multiple myeloma patients who have developed bortezomib resistance as a result of continuous administration of bortezomib to a subject.

  The present invention provides a therapeutic agent for multiple myeloma and a therapeutic agent for bortezomib-resistant multiple myeloma.

FIG. 1 is a graph replacing the drawing in which the cell viability when culturing each cell by adding bortezomib at various concentrations is plotted in a graph. FIG. 2 is a graph instead of a drawing in which the cell viability when culturing each cell by adding plumbagin at various concentrations is plotted in a graph. FIG. 3 is a graph replacing the drawing in which the cell viability when culturing each cell by adding juglone at various concentrations is plotted in a graph. FIG. 4 is a graph instead of a drawing showing the cell viability when LS-102 is administered.

  Hereinafter, modes for carrying out the present invention will be described. The present invention is not limited to the embodiments described below, but includes those appropriately modified by those skilled in the art from the following embodiments.

  The present invention relates to a therapeutic agent for multiple myeloma, and includes an inhibitor of synoviolin activity as an active ingredient. Multiple myeloma is a malignant tumor of plasma cells. Typical clinical symptoms in patients with multiple myeloma are anemia, fractures, hypercalcemia, renal dysfunction, and recurrent infections, which are caused by the growth of malignant tumor cells in the bone marrow.

  An example of a synoviolin activity inhibitor is a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. This compound is known, for example, as described in Table 2014/103862. And it is described in this gazette that this compound is an activity inhibitor of Synoviolin. Synoviolin activity inhibitor means a component that inhibits the activity of Synoviolin. An example of synoviolin activity is ubiquitination. The synoviolin activity inhibitor is a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and these are also referred to as compounds of the present invention.

  In other words, the present invention provides a therapeutic agent for multiple myeloma comprising a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient. provide.

（式（Ｉ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。
Ｘ^１及びＸ^２は，同一でも異なってもよく，酸素原子又は硫黄原子を表す。
Ａ^１−Ａ^２は，Ｃ−Ｃ（単結合），又はＣ＝Ｃ（二重結合）を表す。）

(In the formula (I),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom.
A ¹ -A ² represents C—C (single bond) or C═C (double bond). )

  The above compounds can be isolated and purified using known methods by applying ordinary chemical operations such as extraction, concentration, distillation, crystallization, filtration, recrystallization, and various chromatography. In addition, a single compound may be sufficient as an active ingredient, and multiple types of compounds contained in Formula (I) may be contained. Moreover, when an isomer exists in a compound, only any isomer may be contained as a compound, and a mixture may be sufficient as it.

  The pharmaceutically acceptable salt means a pharmaceutically acceptable salt of the naphthalene derivative represented by the general formula (I). That is, a pharmaceutically acceptable salt means a salt that can be used as a medicine. The pharmaceutically acceptable solvate means a pharmaceutically acceptable solvate of the naphthalene derivative represented by the general formula (I). The same applies to other general formulas. Since these do not greatly change the efficacy of the active ingredient or become the same as the compound in vivo, if the compound has medicinal properties, its salts and solvates can be said to have the same medicinal properties.

  Examples of pharmaceutically acceptable salts are inorganic acid salts, organic acid salts, inorganic base salts, organic base salts, acidic or basic amino acid salts. Examples of inorganic acid salts are hydrochloride, hydrobromide, sulfate, nitrate and phosphate. Examples of organic acid salts are acetate, succinate, fumarate, maleate, tartrate, citrate, lactate, stearate, benzoate, methanesulfonate, and p-toluenesulfone Acid salt. Examples of the inorganic base salt are alkali metal salts such as sodium salt and potassium salt, alkaline earth metal salts such as calcium salt and magnesium salt, aluminum salt, and ammonium salt. Examples of organic base salts are diethylamine salt, diethanolamine salt, meglumine salt, and N, N'-dibenzylethylenediamine salt. Examples of acidic amino acid salts are aspartate and glutamate. Examples of basic amino acid salts are arginine salts, lysine salts, and ornithine salts.

  An example of a solvate is a hydrate.

The C _1-3 alkyl group means an alkyl group having 1 to 3 carbon atoms. Examples of the C _1-3 alkyl group are a methyl group, an ethyl group, and a propyl group (n-propyl group, isopropyl group).

The C _1-3 alkoxy group means an alkoxy group having 1 to 3 carbon atoms. Examples of the C _1-3 alkoxy group are a methoxy group, an ethoxy group, and a propoxy group (n-propoxy group, isopropoxy group).

  Examples of the halogen atom are a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

The compound of the formula (I) preferably has the following substituents.
It is preferable that ^one of R ¹ and R ⁴ is a hydroxyl group and the remaining groups are a hydrogen atom, a hydroxyl group, a methyl group, a methoxy group, or a chlorine atom.
R ² and R ³ are preferably hydrogen atoms.
R ⁵ and R ⁶ are preferably a hydrogen atom or a methyl group, and one of them may be a methyl group and the rest may be a hydrogen atom.

（式（ＩＩ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。
Ｘ^１及びＸ^２は，同一でも異なってもよく，酸素原子又は硫黄原子を表す。） An example of a synoviolin activity inhibitor is a compound represented by the following formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (II),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom. )

The compound of the formula (II) preferably has the following substituents.
It is preferable that ^one of R ¹ and R ⁴ is a hydroxyl group and the remaining groups are a hydrogen atom, a hydroxyl group, a methyl group, a methoxy group, or a chlorine atom.
R ² and R ³ are preferably hydrogen atoms.
R ⁵ and R ⁶ are preferably a hydrogen atom or a methyl group, and one of them may be a methyl group and the rest may be a hydrogen atom.

（式（ＩＩＩ）中，
Ｒ^１〜Ｒ^４は，同一でも異なってもよく，水素原子，水酸基，Ｃ_１−３アルキル基，Ｃ_１−３アルコキシ基，及びハロゲン原子のいずれかを表わす。後述する実施例により実証されたとおり，Ｒ^１〜Ｒ^４の少なくともひとつは水酸基である。
Ｒ^５及びＲ^６は，同一でも異なってもよく，水素原子，Ｃ_１−３アルキル基，及びハロゲン原子のいずれかを表わす。） An example of an activity inhibitor of synoviolin is a compound represented by the following formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (III),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom. )

The compound of the formula (III) preferably has the following substituents.
It is preferable that ^one of R ¹ and R ⁴ is a hydroxyl group and the remaining groups are a hydrogen atom, a hydroxyl group, a methyl group, a methoxy group, or a chlorine atom.
R ² and R ³ are preferably hydrogen atoms.
R ⁵ and R ⁶ are preferably a hydrogen atom or a methyl group, and one of them may be a methyl group and the rest may be a hydrogen atom.

  Specific compounds include 5-hydroxynaphthoquinone (also known as juglone), 5-hydroxy-2-methyl- [1,4] -naphthoquinone (also known as plumbagin), 5,8-dihydroxy-4a, 8a-dihydro- [ 1,4] naphthoquinone, 5-hydroxy-4a, 8a-dihydro- [1,4] naphthoquinone, 5-hydroxy-2,3,4,8a-tetrahydro- [1,4] naphthoquinone, 5-hydroxy-7- Methoxy-4a, 8a-dihydro- [1,4] naphthoquinone, 5-hydroxy-8-methoxy-4a, 8a-dihydro- [1,4] naphthoquinone, and 5-chloro-8-hydroxy-4a, 8a-dihydro -[1,4] naphthoquinone, or two or more thereof.

  Another example of a synoviolin activity inhibitor relates to a 1,3,5-triazine derivative, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. For example, WO 2006-137514 pamphlet describes that a 1,3,5-triazine derivative has a function of controlling self-ubiquitination of synoviolin protein. Similar points are described in Japanese Patent Application Laid-Open No. 2014-141472.

  The 1,3,5-triazine derivative having a synoviolin protein inhibitory activity, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, has the following formula: 1 is a compound represented by 1;

In Formula 1, R ^{11 to} R ¹³ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. R ^{21 to} R ²³ may be the same or different.

Examples of C _1-3 alkyl groups are methyl, ethyl and propyl groups. Examples of the C _1-3 alkoxy group are a methoxy group, an ethoxy group, and a propoxy group. Examples of the halogen atom are a fluorine atom, a chlorine atom, a bromine atom, and an iodine atom.

And when R < ¹¹ > is a hydrogen atom or a halogen atom, R < ²¹ > represents either of the groups represented by Formula 2-6.
On the other hand, when R ¹¹ is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²¹ is any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³¹¹ NHR ³¹² . And may be linked to R ¹¹ to form a ring. -R ³¹¹ represents a C _1-3 alkylene group, and R ³¹² represents a C _1-3 alkyl group.
When R ¹² is a hydrogen atom or a halogen atom, R ²² represents any of the groups represented by formulas 2-6. When R ¹² is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²² is any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³²¹ NHR ³²² . And may be linked to R ¹² to form a ring. -R ³²¹ represents a C _1-3 alkylene group, and R ³²² represents a C _1-3 alkyl group.
When R ¹³ is a hydrogen atom or a halogen atom, R ²³ represents any of the groups represented by formulas 2 to 6. When R ¹³ is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²³ represents any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³³¹ NHR ^332. , R ¹³ may be linked to form a ring. —R ³³¹ represents a C _1-3 alkylene group, and R ³³² represents a C _1-3 alkyl group.

In a preferred embodiment, R ¹¹ and R ¹² represent a hydrogen atom or a halogen atom,
R ¹³ represents a C _1-3 alkyl group or a C _1-3 alkoxy group,
R ²¹ and R ²² may be the same or different and each represents a group represented by formulas 2 to 6;
R ²³ represents —R ³³¹ NHR ³³² ,
—R ³³¹ represents a C _1-3 alkylene group, and R ³³² represents a C _1-3 alkyl group.

In a preferred embodiment, R ¹¹ and R ¹² represent a hydrogen atom,
R ¹³ represents a C _1-3 alkyl group,
R ²¹ and R ²² may be the same or different and each represents a group represented by formulas 2 to 6;
R ²³ represents —R ³³¹ NHR ³³² ,
—R ³³¹ represents a C _1-3 alkylene group, and R ³³² represents a C _1-3 alkyl group.

In a preferred embodiment, R ¹¹ and R ¹² represent a hydrogen atom,
R ¹³ represents a C _1-3 alkyl group,
R ²¹ represents a group represented by Formula 6;
R ²² represents a group represented by Formula 3;
R ²³ represents —R ³³¹ NHR ³³² ,
—R ³³¹ represents a C _1-3 alkylene group, and R ³³² represents a C _1-3 alkyl group.

Preferred as 1,3,5-triazine derivatives having inhibitory activity of noviolin protein, 1,3,5-triazine derivatives thereof, or pharmaceutically acceptable salts thereof, or pharmaceutically acceptable solvates thereof , A substance (LS-102) having the following chemical formula. This substance is a synoviolin activity inhibitor as described in, for example, Japanese Patent No. 6208689. Compound name LS-102 is, (S) -N ² - (benzo [d] oxazole --6-yl) -N ⁴ - (cyclohexylethyl) -N ⁶ - ethyl -N ⁶ - (2- (ethyl amino) ethyl) -1,3,5-triazine-2,4,6-triamine ^{((S) -N 2 - (} benzo [d] oxazol-6-yl) -N 4 - (1-cyclohexylethyl) -N ^{6 -ethyl-N 6} - is (2- (ethylamino) ethyl) -1,3,5 -triazine-2,4,6-triamine).

  The therapeutic agent for multiple myeloma of the present invention can be preferably used as a therapeutic agent for bortezomib-resistant multiple myeloma. For example, this therapeutic agent is preferably used as a therapeutic agent for multiple myeloma for multiple myeloma patients who have developed bortezomib resistance as a result of continuous administration of bortezomib to a subject.

  The drug of the present invention can be mixed with a physiologically acceptable carrier, excipient, diluent or the like and administered orally or parenterally as a pharmaceutical composition. As oral preparations, dosage forms such as granules, powders, tablets, capsules, solvents, emulsions or suspensions can be used. As the parenteral preparation, a dosage form such as an injection, an infusion, an external medicine, or a suppository can be selected. Examples of injections include subcutaneous injections, intramuscular injections, intravenous injections, and intraperitoneal injections. As an external medicine, a nasal administration agent, an inhalant (a spray), an ointment, or the like can be shown. The preparation technology for the above dosage form so as to include the drug of the present invention as the main component is known. For example, a tablet for oral administration can be produced by adding an excipient, a disintegrant, a binder, a lubricant, and the like to the drug of the present invention, and mixing and compression-molding. As the excipient, lactose, starch, mannitol or the like is generally used. As the disintegrant, calcium carbonate or carboxymethyl cellulose calcium is generally used. As the binder, gum arabic, carboxymethylcellulose, or polyvinylpyrrolidone is used. Known lubricants include talc and magnesium stearate.

  An injection can be obtained by dissolving the drug of the present invention as a main component together with an appropriate dispersant, dissolving in a dispersion medium, or dispersing. Depending on the choice of the dispersion medium, either aqueous or oily solvent dosage forms can be used. In order to use an aqueous solvent, distilled water, physiological saline, Ringer's solution, or the like is used as a dispersion medium. For oil-based solvents, various vegetable oils and propylene glycol are used as dispersion media. At this time, a preservative such as paraben may be added as necessary. In addition, known isotonic agents such as sodium chloride and glucose can be added to the injection. In addition, soothing agents such as benzalkonium chloride and procaine hydrochloride can be added.

  Moreover, it can be set as an external preparation by making the chemical | medical agent of this invention into a solid, liquid, or semi-solid composition. About a solid or liquid composition, it can be set as an external preparation by setting it as the composition similar to what was described previously. A semi-solid composition can be prepared by adding a thickener to an appropriate solvent as required. As the solvent, water, ethyl alcohol, polyethylene glycol, or the like can be used. As the thickener, bentonite, polyvinyl alcohol, acrylic acid, methacrylic acid, polyvinylpyrrolidone, or the like is generally used. A preservative such as benzalkonium chloride can be added to the composition. A suppository can also be obtained by combining an oily base material such as cacao butter or an aqueous gel base material such as a cellulose derivative as a carrier.

  The necessary amount (effective amount) of the drug of the present invention is administered to mammals including humans within the safe dose range. The dosage of the drug of the present invention can be appropriately determined finally by the judgment of a doctor or veterinarian in consideration of the type of dosage form, administration method, patient age and weight, patient symptoms, and the like. The agent of the present invention is administered to a subject once to several times per day, for example. When the drug of the present invention is administered to a subject, the compound of the present invention, which is an active ingredient, may be administered, for example, at 1 μg or more and 10 g or less at a time, and may be administered at 10 μg or more and 100 mg or less. Furthermore, 1 mg or more and 25 mg or less may be administered per dose.

  The present invention also provides a method for treating multiple myeloma, comprising the step of administering an effective amount of a synoviolin activity inhibitor (or a compound of the present invention) to a target human or non-human mammal.

Mechanism of resistance to bortezomib Over time, multiple myeloma that develops resistance to bortezomib appears. The primary mechanism for acquiring resistance is that the proteasome acquires mutations and will not accept inhibition by bortezomib. The absence of inhibition by bortezomib removes ubiquitinated denatured proteins and allows multiple myeloma to survive. On the other hand, inhibition of Synoviolin does not cause ubiquitination of the denatured protein itself, and can induce cell death in multiple myeloma that has acquired resistance to bortezomib.

[Example 1]
Four types of cells: A549 cell as a general cancer cell line, HeLa cell, KMS11 cell as a cancer cell line for multiple myeloma, and KMS11-BTZ cell as a multiple myeloma cancer cell line that has acquired resistance to bortezomib Prepared. A549 cells were cultured in DMEM medium containing 10% FCS, HeLa cells were cultured in MEM medium containing 10% FCS, KMS11 cells and KMS11-BTZ details were cultured in RPMI 1640 medium containing 10% FCS.
In order to measure the cytotoxicity of bortezomib, plumbagin and juglone, cells were seeded in a 48-well plate. HeLa and A549 were seeded by suspending 1.25 × 10 ⁴ cells in 200 μl medium, and KMS11 and KMS11-BTZ were seeded by suspending 2.5 × 10 ⁴ cells in 200 μl medium. Simultaneously with cell seeding, bortezomib, plumbagin and juglone were added at various concentrations and cultured in the presence of 37 ° C. and 5% CO ₂ . After culturing for 3 days, 10 μl of cell counting kit 8 (Dojindo Laboratories) was added to each well and reacted at 37 ° C. in the presence of 5% CO ₂ for 1 hour. Absorbance was measured. The cell viability was calculated from the absorbance value when the value when no drug was added was 1. The IC ₅₀ value for each cell of each drug was calculated from the obtained cell viability.

The results are shown in FIGS. FIG. 1 is a graph replacing the drawing in which the cell viability when culturing each cell by adding bortezomib at various concentrations is plotted in a graph. The cell survival rate when no drug is added is set to 1. IC ₅₀ values for each cell are also shown. The horizontal axis indicates the concentration (nM) of bortezomib. From the lowest IC ₅₀ value, KMS11, A549, Hela and KMS11-BTZ are in this order.
FIG. 2 is a graph instead of a drawing in which the cell viability when culturing each cell by adding plumbagin at various concentrations is plotted in a graph. The cell survival rate when no drug is added is set to 1. IC ₅₀ values for each cell are also shown. The horizontal axis is the plumbagin concentration (nM). From the lowest IC ₅₀ value, KMS11-BTZ, KMS11, Hela, and A549 are in this order.
FIG. 3 is a graph replacing the drawing in which the cell viability when each cell is cultured with juglone added at various concentrations is plotted in a graph. The cell survival rate when no drug is added is set to 1. The horizontal axis indicates the concentration (nM) of juglone. IC ₅₀ values for each cell are also shown. From the lowest IC ₅₀ value, KMS11-BTZ, KMS11, Hela, and A549 are in this order.

  Bortezomib showed higher cytotoxicity to KMS11 cells than to general cancer cells such as A549 cells and HeLa cells. On the other hand, it did not show cytotoxicity against KMS11-BTZ, which is a bortezomib resistant cell.

  Synoviolin inhibitors plumbagin and juglone showed higher cytotoxicity against KMS11 cells than to general cancer cells such as A549 cells and HeLa cells. As with KMS11 cells, KMS11-BTZ, which is a bortezomib-resistant cell, showed high cytotoxicity.

Discussion The Synoviolin Activity Inhibitor is Effective as a Treatment Agent for Multiple Myeloma Hereinafter, based on Examples, the mechanism of action in which a Synoviolin activity inhibitor is effective as a treatment agent for multiple myeloma will be discussed.
Various proteins are synthesized in the cell. A protein is a series of 20 amino acids. And this long chain of amino acids can be folded in a more complicated way to obtain various functions. In the process of producing a protein, a protein that does not fold correctly is also created (this is called a denatured protein). Accumulation of denatured proteins in the cell results in endoplasmic reticulum stress and ultimately cell death. Therefore, the denatured protein needs to be removed from the cell. When the denatured protein is removed, the denatured protein is first modified by ubiquitination by the action of an enzyme called Synoviolin. Next, the ubiquitinated denatured protein is degraded by a cellular structure called a proteasome.

  Bortezomib (trade name: Velcade), which is approved for multiple myeloma, is a molecularly targeted drug that inhibits the action of the proteasome. In multiple myeloma, antibody-producing cells in the bone marrow are tumorized. Therefore, protein synthesis is more active than other cells, and it is probable that a denatured protein is probable. Therefore, the accumulation of denatured protein by bortezomib and cell death occur preferentially in multiple myeloma.

  As mentioned above, Synoviolin is an enzyme that ubiquitinates denatured proteins. By inhibiting synoviolin, the denatured protein escapes proteasome degradation and accumulates. As a result, cell death in multiple myeloma may be preferentially caused, as is proteasome inhibition by bortezomib.

[Example 2]
Next, it was examined whether LS-102, which is a selective inhibitor of Synoviolin's E3 ubiquitin ligase activity, has cytotoxic activity specific to multiple myeloma.
Three types of cells, HeLa cells, KMS11 cells, and bortezomib-resistant KMS11-BTZ cells were added to 5 μM LS-102 and cultured.
After culturing at 37 ° C. in the presence of 5% CO ₂ for 3 days, 10 μl of cell counting kit 8 (Dojindo Laboratories) was added to each well, and at 37 ° C. in the presence of 5% CO ₂ for 1 hour. After the reaction, the absorbance at 450 nm was measured with a microplate reader. The cell viability was calculated from the absorbance value when the value when no drug was added was 1. The result is shown in FIG. FIG. 4 is a graph instead of a drawing showing the cell viability when LS-102 is administered.

  Since the present invention relates to a therapeutic agent for multiple myeloma, it can be used in the pharmaceutical industry.

Claims (7)

  A therapeutic agent for multiple myeloma containing an inhibitor of synoviolin activity as an active ingredient. The therapeutic agent for multiple myeloma according to claim 1,
A therapeutic agent for multiple myeloma, wherein the synoviolin activity inhibitor is a compound represented by the following formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (I),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom.
A ¹ -A ² represents C—C (single bond) or C═C (double bond). ) The therapeutic agent for multiple myeloma according to claim 1,
A therapeutic agent for multiple myeloma, wherein the synoviolin activity inhibitor is a compound represented by the following formula (II), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (II),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom.
X ¹ and X ² may be the same or different and each represents an oxygen atom or a sulfur atom. ) The therapeutic agent for multiple myeloma according to claim 1,
A therapeutic agent for multiple myeloma, wherein the synoviolin activity inhibitor is a compound represented by the following formula (III), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

(In the formula (III),
R ^{1 to} R ⁴ may be the same or different and each represents a hydrogen atom, a hydroxyl group, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom. As demonstrated by the examples described later, at least one of R ^{1 to} R ⁴ is a hydroxyl group.
R ⁵ and R ⁶ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, or a halogen atom. ) The therapeutic agent for multiple myeloma according to claim 1,
A therapeutic agent for multiple myeloma, wherein the synoviolin activity inhibitor is a compound represented by the following formula 1, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof.

In Equation 1,
R ^{11 to} R ¹³ may be the same or different and each represents a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, or a halogen atom;
R ^{21 to} R ²³ may be the same or different,
When R ¹¹ is a hydrogen atom or a halogen atom, R ²¹ represents any of the groups represented by formulas 2 to 6;
When R ¹¹ is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²¹ represents any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³¹¹ NHR ^312. , R ¹¹ may be linked to form a ring,
—R ³¹¹ represents a C _1-3 alkylene group, R ³¹² represents a C _1-3 alkyl group,

When R ¹² is a hydrogen atom or a halogen atom, R ²² represents any of the groups represented by formulas 2 to 6;
When R ¹² is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²² represents any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³²¹ NHR ^322. , R ¹² may be linked to form a ring,
—R ³²¹ represents a C _1-3 alkylene group, R ³²² represents a C _1-3 alkyl group,

When R ¹³ is a hydrogen atom or a halogen atom, R ²³ represents any of the groups represented by formulas 2 to 6;
When R ¹³ is a C _1-3 alkyl group or a C _1-3 alkoxy group, R ²³ represents any one of a hydrogen atom, a C _1-3 alkyl group, a C _1-3 alkoxy group, and —R ³³¹ NHR ^332. , R ¹³ may be linked to form a ring,
—R ³³¹ represents a C _1-3 alkylene group, and R ³³² represents a C _1-3 alkyl group.

The therapeutic agent for multiple myeloma according to claim 1,
The therapeutic agent for multiple myeloma is a therapeutic agent for bortezomib-resistant multiple myeloma,
A therapeutic agent for multiple myeloma. The therapeutic agent for multiple myeloma according to claim 1,
The therapeutic agent for multiple myeloma is a therapeutic agent for multiple myeloma for a patient with multiple myeloma that has developed resistance to bortezomib as a result of continuous administration to bortezomib.
A therapeutic agent for multiple myeloma.

Images