WO2020209379A1 - Pharmaceutical composition for administration to pregnant women or women who may be pregnant - Google Patents

Abstract

The purpose of the invention is to develop a pharmaceutical product that can be used during pregnancy. Inventors have found that placental permeability can be controlled by controlling, in particles having on the surface thereof a hydrophilic polymer to which a therapeutic drug is bonded or coordinated, the particle size of the particles by the hydrophilic polymer. The problem is solved by particles having a particle size of 10-100 nm when measured by dynamic light scattering.

Description

Pharmaceutical composition for administration to pregnant women or women of childbearing potential

The present invention relates to a pharmaceutical composition for administration to a pregnant woman or a woman who may become pregnant.

Medications for pregnant women are avoided as much as possible to avoid affecting the fetus. Effects on the fetus include teratogenicity, miscarriage, premature birth and fetal stunting. The number of drugs for which safety has been established is limited, and it is necessary to discontinue medication when pregnancy is known. Pregnant women may also suffer from excessive labor due to the discovery of pregnancy during medication. On the other hand, discontinuation of medication often has an adverse effect on pregnant women, and development of a drug that can be used even during pregnancy is desired. Cancer is one of the diseases that becomes a problem during pregnancy. Approximately 200,000 pregnant women worldwide suffer from cancer, of which breast cancer has been reported to be associated with exposure to endogenous estrogen and progesterone and develops with pregnancy. There is also. The risk of breast cancer increases with age, but the number of cases of breast cancer in pregnant women increases with increasing age of pregnancy. Generally, for breast cancer, anticancer agents, hormone therapy, antibody therapeutic agents, radiation therapy, surgery, etc. are performed, but the anticancer agents that can be used for pregnant women are limited.

Inflammation during pregnancy, especially bacterial inflammation, is known to increase the risk of preterm birth, and non-steroidal anti-inflammatory drugs such as indomethacin can be used for treatment. On the other hand, it has been reported that non-steroidal anti-inflammatory drugs such as indomethacin affect the fetus and cause renal failure and digestive failure, and sufficient caution is required for their use.

Preeclampsia is known as a disease that occurs frequently in pregnant women, and it occurs in about 1 in 20 people. Preeclampsia impairs renal and hepatic function, is dangerous to the mother, and can also cause fetal growth restriction. Antihypertensive agents such as hydralazine, methyldopa, labetalol, and nifedipine are commonly used to treat preeclampsia. Recent studies have reported that statins are useful for the treatment and / or prevention of preeclampsia (Non-Patent Document 1: Plos One (2010) vol. 5, Issue 10, e13663). The use of statins in pregnant women is contraindicated. In addition, some psychiatric drugs administered for the treatment of psychiatric disorders may cause teratogenicity or neonatal maladaptation syndrome, and the medication may be discontinued.

International Publication No. 96/32343 International Publication No. 96/33233 International Publication No. 97/06202

The purpose is to develop a drug that can be used during pregnancy.

The present inventors conducted research focusing on the placental permeability of the drug for the purpose of developing a drug that can be used during pregnancy. We have found that placental permeability can be controlled by controlling the size with a hydrophilic polymer, and have reached the present invention. Therefore, the present invention relates to the following:

[1] A pharmaceutical composition for administration to a pregnant woman or a woman of childbearing potential, which comprises particles having a hydrophilic polymer on the surface to which a therapeutic agent is bound or coordinated, and has a dynamic particle size. The pharmaceutical composition having a diameter of 10 to 100 nm as measured by a light scattering method.
[2] The pharmaceutical composition according to item 1, wherein the particles are micelles containing a polyethylene glycol-polyamino acid block copolymer.
[3] The polyethylene glycol-polyamino acid block copolymer has the following general formula (I) or (II):

(During the ceremony
R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
R ^2a , and R ^2c , independently on each appearance, either do not exist or represent a methylene group.
R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
m is an integer from 20 to 2,000
n is an integer from 1 to 200)
Item 2. The block copolymer represented by, wherein the therapeutic agent is coordinated via R ^2b and R ^2d of the block copolymer, or is bound directly or via a linker. The pharmaceutical composition described.
[4] The pharmaceutical composition according to any one of items 1 to 3, wherein the particle size is 25 to 75 nm.
[5] The pharmaceutical composition according to any one of items 1 to 4, wherein the therapeutic agent is administered at a dosage and / or dosage that is contraindicated for pregnant women when administered only with the therapeutic agent.
[6] The pharmaceutical composition according to any one of items 1 to 5, wherein the therapeutic agent is selected from the group consisting of an anticancer drug, an anti-inflammatory drug, an antihypertensive drug, and a psychotropic drug.
[7] The pharmaceutical composition according to item 6, wherein the anticancer drug is a platinum preparation.
[8] The pharmaceutical composition according to item 7, wherein the platinum preparation is formed through a coordination bond with one or two carboxylate groups of the block copolymer.
[9] The pharmaceutical composition according to any one of items 1 to 6, wherein the therapeutic agent is an anti-inflammatory agent and is used for preventing miscarriage.
[10] The pharmaceutical composition according to item 9, wherein the anti-inflammatory drug is a non-steroidal anti-inflammatory drug.
[11] The pharmaceutical composition according to any one of items 1 to 6, wherein the therapeutic agent is an antihypertensive agent and is for the treatment of a gestational hypertension agent.
[12] The pharmaceutical composition according to item 11, wherein the antihypertensive drug is a statin-based antihypertensive drug.
[13] The pharmaceutical composition according to any one of items 1 to 12, wherein the pharmaceutical composition is a pharmaceutical composition for reducing fetal phytotoxicity.
[14] The pharmaceutical composition according to any one of items 1 to 12, wherein the pharmaceutical composition is a pharmaceutical composition for reducing placental barrier permeation.
[15] The following general formula (I) or (II):

(During the ceremony
R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
R ^2a and R ^2c independently exist for each occurrence or represent a methylene group.
R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
m is an integer from 20 to 2,000
n is an integer from 1 to 200)
A drug is a drug-combined block copolymer in which a drug is bound to the block copolymer represented by (1) via R ^2b and R ^2d of the block copolymer, directly or via a linker, and the drug is , Indomethacin or simvastatin, said drug complex block copolymer.
[16] The pharmaceutical composition containing micelles formed from the sound drug complex block copolymer according to item 15, wherein the particle size is 20 to 100 nm when measured by a dynamic light scattering method. Pharmaceutical composition.
[17] The pharmaceutical composition according to item 16, wherein the drug contains indomethacin, and the pharmaceutical composition is a miscarriage-preventing pharmaceutical composition.
[18] The pharmaceutical composition according to item 16, wherein the drug contains simvastatin, and the pharmaceutical composition is a pharmaceutical composition for the treatment of preeclampsia.
[19] A method for producing micelles, which comprises a polyethylene glycol-polyamino acid block copolymer to which a therapeutic agent is bound or coordinated and is to be administered to a pregnant woman or a woman of childbearing potential.
A step of reconstitution of the polyethylene glycol-polyamino acid block copolymer in water, and
A step of obtaining micelles having a particle size of 20 to 100 nm by performing ultrafiltration using a dialysis membrane having a molecular weight cut off of 10,000 to 100,000.
The production method.
[20] The production method according to item 19, wherein the therapeutic agent is selected from the group consisting of an anticancer drug, an anti-inflammatory drug, an antihypertensive drug, and a psychotropic drug.
[21] The polyethylene glycol-polyamino acid block copolymer has the following general formula (1-a) or (1-b):

(During the ceremony
R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
R ^2a and R ^2c independently exist for each occurrence or represent a methylene group.
R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue.
Here, a is an integer of 1 to 5, and X is an amine compound residue containing one or more of primary, secondary, tertiary amines or quaternary ammonium salts, or a compound that is not an amine. Is a residue
m is an integer from 20 to 2,000
n is an integer from 1 to 200)
Item 19 or item 19 or the block copolymer represented by, wherein the therapeutic agent is coordinated via R ^2b or R ^2d of the block copolymer, or is bound directly or via a linker. The manufacturing method according to 20.
[22] Particles having a hydrophilic polymer on the surface to which a therapeutic agent for use in the treatment or prevention of a disease in a pregnant woman or a woman of childbearing potential is bound or coordinated, and the particle size is dynamic light. The particles having a diameter of 10 to 100 nm as measured by a scattering method.
[23] Particles having a hydrophilic polymer on the surface to which an anti-inflammatory drug for use in the prevention, prevention or treatment of miscarriage or premature birth is bound or coordinated in a pregnant woman or a woman of childbearing potential. The particles having a diameter of 10 to 100 nm as measured by a dynamic light scattering method.
[24] In pregnant women or women of childbearing potential, particles having a hydrophilic polymer on the surface to which an antihypertensive drug for use in the prevention or treatment of preeclampsia is bound or coordinated, and having a particle size of The particles having a diameter of 10 to 100 nm as measured by a dynamic light scattering method.
[25] The particle according to any one of items 22 to 24, which reduces fetal phytotoxicity.
[26] The particle according to any one of items 22 to 24, wherein the placental barrier permeability is reduced.
[27] A method of treating or preventing a disease in a pregnant woman or a woman of childbearing potential, in which particles having a hydrophilic polymer on the surface to which a therapeutic agent is bound or coordinated to the pregnant woman or woman of childbearing potential. The method, wherein the particle size of the particles is 10 to 100 nm as measured by a dynamic light scattering method.
[28] In pregnant women or women of childbearing potential, a method for preventing, preventing or treating miscarriage or premature birth, in which particles having a hydrophilic polymer on the surface to which an anti-inflammatory agent is bound or coordinated are provided. The method described above, comprising administering to a pregnant woman or a woman of childbearing potential, wherein the particle size is 10-100 nm as measured by dynamic light scattering.
[29] In a pregnant woman or a woman of childbearing potential, a method for preventing or treating preeclampsia, in which particles having a hydrophilic polymer on the surface to which an antihypertensive drug is bound or coordinated are provided on the surface of the pregnant woman or pregnant woman. The method described above, comprising administering to a potential woman, wherein the particle size is 10-100 nm as measured by dynamic light scattering.
[30] The method according to any one of items 27 to 29, wherein the phytotoxicity of the fetus is reduced.
[31] The method according to any one of items 27-29, wherein the placental barrier permeability is reduced.
[32] The surface has a hydrophilic polymer to which a therapeutic agent is bound or coordinated and has a particle size for the manufacture of a medicament for use in the treatment or prevention of a disease in a pregnant woman or a woman of childbearing potential. Use of particles that are 10-100 nm as measured by dynamic light scattering.
[33] In pregnant women or women of childbearing potential, particles to which a therapeutic agent is bound or coordinated to produce a drug for the prevention, prevention or treatment of miscarriage or preterm birth, the hydrophilic polymer. Use of particles on the surface that have a particle size of 10-100 nm as measured by dynamic light scattering.
[34] In pregnant women or women of childbearing potential, particles to which a therapeutic agent is bound or coordinated for the manufacture of a pharmaceutical for the prevention or treatment of preeclampsia, with a hydrophilic polymer on the surface. However, the use of particles whose particle size is 10 to 100 nm when measured by dynamic light scattering.
[35] The use according to any one of items 32 to 34, wherein the fetal phytotoxicity is reduced.
[36] The use according to any one of items 32 to 34, wherein the placental barrier permeability is reduced.

The pharmaceutical composition of the present invention has low placental permeability and can reduce the effect of the drug on the fetus. This makes it possible to provide a medicine that can be administered to a pregnant woman or a woman who may become pregnant.

FIG. 1 is a photograph showing the accumulation of high molecular weight micelles in the placenta and fetus. FIG. 2 is a photograph showing the analysis of elements (Pt, Fe, K, Ca) after administration of micelles having a size of 30 nm. FIG. 3 is a graph showing the platinum content in the fetus of pregnant mice treated with oxaliplatin and dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm). FIG. 4 is a graph showing the platinum content in the placenta of pregnant mice treated with oxaliplatin and dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm). FIG. 5 is a graph showing placental permeability of a drug by a human placental perfusion model. FIG. 5A shows the results for oxaliplatin, 30 nm dahaplatin-encapsulating micelles, 70 nm dahaplatin-encapsulating micelles, and 8-armPEG. FIG. 5B shows the results for 10 nm, 20 nm, and 30 nm PEG-coated gold nanoparticles. FIG. 6 is a graph showing the amount of drug accumulated in the human placenta. FIG. 7 shows the ¹ H-NMR spectrum of PEG-poly (Asp-Furan-OH). FIG. 8 is a graph showing the stability of micelles in blood and endosome pH. FIG. 9 is a graph showing the amount of indomethacin released in macrophages. FIG. 10 is a graph showing placental permeability of a drug by a human placental perfusion model. FIG. 11 is a diagram showing the tissue transferability of indomethacin-encapsulating polymer micelles. FIG. 11A shows the location of the recovered organs, and FIGS. 11B-E show the fluorescence of indomethacin-encapsulating polymer micelles at 1, 4, 8 and 24 hours after administration. These experiments show that indomethacin micelles are distributed in the liver, kidneys, spleen, uterine body, and placenta, and do not reach the fetus at each point in time. FIG. 12 is a diagram showing an evaluation of tissue migration of indomethacin-encapsulating polymer micelles. The amount of indomethacin transferred to (A) kidney, (B) liver, (C) spleen, (D) placenta, and (E) fetus after administration of indomethacin-encapsulating polymer micelles to pregnant mice is shown. It is a graph which shows the survival rate by administration of indomethacin, indomethacin-encapsulating polymer micelle in a preterm mouse model. FIG. 14 is a graph showing changes in blood pressure after administration of a drug in a preeclampsia model mouse. FIG. 15 is a graph showing changes in fetal body weight after administration of a drug in a preeclampsia model mouse. FIG. 16 is a graph showing the amount of urinary albumin after drug administration in a preeclampsia model mouse.

The present invention relates to a pharmaceutical composition for administration to a pregnant woman or a woman who may become pregnant. The pharmaceutical composition of the present invention contains particles having a hydrophilic polymer to which a therapeutic agent is bound or coordinated on the surface, and the particle size thereof is 10 to 100 nm when measured by a dynamic light scattering method. It is a feature. From the viewpoint of placental permeability, the particle size is usually 10 nm or more, preferably 20 nm or more, and more preferably 25 nm or more. From the viewpoint of exerting the efficacy of the therapeutic agent, the particle size is usually 100 nm or less, preferably 90 nm or less, more preferably 80 nm or less, and even more preferably 75 nm or less. Particles of this size have low placental permeability and can reduce fetal toxicity. In the composition of the present invention, the polydispersity (PDI) of the particle size of the particles is preferably 0.2 or less, and even more preferably 0.1 or less. Since the particles contained in the pharmaceutical composition of the present invention have reduced placental permeability, they can be used as a pharmaceutical composition for reducing fetal phytotoxicity or reducing placental barrier penetration.

Any particles may be used as long as the particles have a hydrophilic polymer on the surface, and may be micelles or vesicles formed by a block copolymer, liposomes whose surface is covered with a hydrophilic polymer, or the like.

As the hydrophilic polymer, any polymer can be used as long as it is a hydrophilic polymer generally used in medicines, particularly drug delivery systems. As an example, polyethylene glycol, polylactic acid, polyglycolic acid, polypeptide, polysaccharide and the like can be mentioned. From the viewpoint of producing micelles, polyethylene glycol-polyamino acid block copolymers can be used. As the polyethylene glycol-polyamino acid block copolymer, any known in the art can be used. As the amino acid of the polyethylene glycol-polyamino acid block copolymer, glutamic acid or aspartic acid having a carboxy group in the side chain can be used. As an example, the following general formula (I) or (II):

(During the ceremony
R ^1a and R ^1b represent a hydrogen atom, a hydroxyl group, or an unsubstituted or substituted linear or branched C _1-12 alkyl group or C _1-12 alkoxy group.
L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
R ^2a and R ^2c independently exist on each occurrence or represent a methylene group.
R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
m is an integer from 20 to 2,000
n is an integer from 1 to 200)
Block copolymers represented by can be used. Therapeutic agents are bound or coordinated in R ^2b and R ^2d either directly or via a linker.

Here, in the structural formulas of the general formulas (I) and (II), when the segment having the number of repeating units (degree of polymerization) of "m" is displayed as a PEG-derived uncharged hydrophilic segment (hereinafter referred to as "PEG segment"). The segment whose number of repeating units is "n" is an amino acid-derived segment (hereinafter, may be referred to as "polyamino acid segment").

In the general formulas (I) and (II), R ^1a and R ^1b are independently hydrogen atoms, hydroxyl groups, or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _{1-12, respectively.} Represents an alkoxy group. Linear or branched C _1-12 include, for example, methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl, decyl, undecyl. And so on. When substituted, the substituents include acetalized formyl group, cyano group, formyl group, carboxyl group, amino group, C _1-6 alkoxycarbonyl group, C _2-7 acylamide group, and the same or different tri-C _{1 -6} Alkylsiloxy group, siloxy group or silylamino group can be mentioned. Here, acetalization refers to the reaction of the carbonyl of formyl with, for example, two molecules of an alkanol having 1 to 6 carbon atoms or a branched alkylene diol having 2 to 6 carbon atoms. It means formation and is also a method of protecting the carbonyl group. For example, when the substituent is an acetalized formyl group, it can be hydrolyzed under mild acidic conditions and converted to another substituent, a formyl group (-CHO) (or aldehyde group).

In the general formulas (I) and (II), L ¹ and L ² represent a linking group. Specifically, L ¹ is preferably − (CH ₂ ) _b −NH− (where b is an integer of 1 to 5), and L ² is − (CH ₂ ) _c −CO− (here). And c is an integer of 1 to 5).

In the general formulas (I) and (II), R ^2a and R ^2c do not exist independently at each appearance or represent a methylene group. When aspartic acid is used as the amino acid, dehydration condensation occurs between the carboxy group in the side chain of aspartic acid and the nitrogen in the main chain, and a succinimide intermediate is produced. Cleavage of this intermediate can then result in isomerization to isoaspartic acid. Therefore, when aspartic acid is isomerized to isoaspartic acid, a methylene group is generated at the positions of R ^2a and R ^2c , and R ^2b and R ^2d become carboxy groups. In the absence of R ^2a and R ^2c, isomerization does not occur, indicating that the amino acid has a peptide bond in the main chain. The rate of isomerization in the block copolymer can be arbitrary. Thus, in the absence of R ^2a and R ^2c , R ^2b and R ^2d represent any amino acid side and the drug is coordinated through these groups or the drug, either directly or via a linker. Can be combined. The amino acid side chain may be a side chain of any biological constituent amino acid, but from the viewpoint of binding a drug, it may be a side chain of serine, threonine, arginine, histidine, lysine, cysteine, glutamic acid and aspartic acid. preferable. When R ^2a and R ^2c are present, R ^2b and R ^2d are carboxy groups. Since the isomerization of aspartic acid to isoaspartic acid in the polyamino acid segment can occur accidentally, the above description is made even when the formula representing only the amide bond between the main chains is described in the present specification. It is not intended to exclude isomerized polymers as in formulas (I) and (II) of. Repetitive units of amino acids can be present in blocks, or can be randomly present.

In the general formulas (I) and (II), R ³ represents a hydrogen atom, a protecting group, a hydrophobic group or a polymerizable group. Specifically, R ³ is preferably an acetyl group, an acryloyl group, or a methacryloyl group.

In the general formulas (I) and (II), R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue. Here, a is an integer of 1 to 5, and X is an amine compound residue containing one or more of primary, secondary, tertiary amines or quaternary ammonium salts, or a compound that is not an amine. It is preferably a residue.

In the general formulas (I) and (II), m is an integer of 20 to 2,000, preferably an integer of 40 to 500, and more preferably an integer of 45 to 300. Further, n is an integer of 1 to 200, preferably an integer of 20 to 100, and more preferably an integer of 30 to 50.

Each repeating unit in the general formulas (I) and (II) is shown in the order specified for convenience of description, but each repeating unit can exist in a random order. In particular, it is preferred that only each repeating unit in the polyamino acid segment can be present in random order as described above.

The molecular weight (Mw) of the block copolymer represented by the general formulas (I) and (II) is not limited, but is preferably 5,000 to 40,000, more preferably 8,000 to 22,000. is there. For each segment, the molecular weight (Mw) of the PEG segment is preferably 1,000 to 20,000, more preferably 2,000 to 12,000, and the molecular weight of the polyamino acid segment (Mw). ) Is preferably 4,000 to 20,000 as a whole, and more preferably 6,000 to 10,000.

Method for producing a block copolymer represented by the general formula (I) and (II) include, but are not limited to, for example, R ^1a - or R ^1b - and previously synthesized segment (PEG segment) comprising a block portion of the PEG chain ; then, one end of the PEG segment (R ^1a - - or R ^1b opposite ends), and polymerizing a prescribed monomers in sequence, substitution or to include an anionic group to the side chain then optionally Examples thereof include a method of conversion, a method of synthesizing the PEG segment and a block portion having a side chain containing an anionic group in advance, and connecting them to each other. The methods and conditions of various reactions in the production method can be appropriately selected or set in consideration of the conventional method. The PEG segment is a method for producing a PEG segment portion of a block copolymer described in, for example, Patent Document 1: WO96 / 32434, Patent Document 2: WO96 / 33233, and Patent Document 3: WO97 / 06202. Can be prepared using.

The bond between the PEG segment portion and the polyamino acid segment portion thus formed can also take an appropriate linking mode depending on the method for producing the block copolymer represented by the general formula (I) or (II), and the present invention As long as it meets the object of the invention, it may be bonded by any linking group. The production method is not particularly limited, but as one method for producing the polymer of the general formula (I) or (II), a PEG derivative having an amino group at the terminal is used, for example, from the amino terminal thereof. , Β-benzyl-L-aspartate (BLA), γ-benzyl-L-glutamate and other protected amino acids N-carboxylic acid anhydrides (NCA) are ring-open polymerized to synthesize block copolymers, followed by Examples thereof include a method of converting a chain benzyl group into another ester group, or partially or completely hydrolyzing the chain benzyl group to obtain the desired block copolymer. In this case, the structure of the copolymer is the general formula (I), and the linking group L1 is a structure derived from the terminal structure of the PEG segment used, but is preferably − (CH ₂ ) _b −NH− (here). , B is an integer from 1 to 5).

Further, the copolymer of the present invention can also be produced by a method of synthesizing a polyamino acid segment portion and then binding it to a PEG segment portion prepared in advance, and in this case, the result is the same as that produced by the above method. However, the linking group L ₂ is not particularly limited, but is preferably − (CH ₂ ) _c −CO−, as it may have a structure corresponding to the general formula (II). Yes (where c is an integer from 1 to 5).

Drugs can be bound or coordinated to the hydrophilic polymers of the present invention. The agent can be attached to or coordinated with the hydrophilic polymer via a linker or directly. When the hydrophilic polymer is a block copolymer of formula (I) or (II), the agent can be coordinated or attached at R ^2b and R ^2d . When the drug contains a heavy metal such as platinum, the carboxylate group formed by liberating the proton of the carboxy group of the amino acid side chain coordinates with the heavy metal of the drug. When the drug is directly bound, it can be bound via the carboxy group, amino group, thiol group of the amino acid side chain. Any group of the drug can be used for binding, but carboxy groups, amino groups, thiol groups and the like can be used as an example. When attached via a linker, the linker L ₃ can be any linker used in the art, such as C _1-6 alkylene. In yet another embodiment, a drug in which a hydrazine is reacted with a carboxy group of a polyamino acid or a derivative thereof to form a hydrazide group (in this case, M is -NH-NH ₂ ) and has a ketone via the hydrazide group. May be combined with. In this case, the linker L becomes a hydrazone bond. As the linker that binds the drug to the polyamino acid, any linker known in the art can be used from the viewpoint of producing desired particles.

The therapeutic agent contained in the pharmaceutical composition of the present invention may be any therapeutic agent. Particularly in pregnant women, the required therapeutic agents are mentioned, and as an example, anticancer agents, anti-inflammatory agents, antihypertensive agents, psychotropic agents and the like can be used. Examples of anticancer agents include anticancer agents for treating cancer in any organ such as stomach, skin, larynx, mouth, pharynx, esophagus, digestive organs, pancreas, lung, brain, bone, bone marrow, and breast. Drugs for the treatment of breast cancer, which are more likely to occur in pregnant women.

As the drug used as an anticancer drug, a platinum preparation, an alkylating drug, an antimetabolite, a topoisomerase inhibitor, a microtubule inhibitor, an endocrine therapeutic drug, etc. can be used. Platinum preparations include dahaplatin, oxaliplatin, cisplatin, carboplatin and nedaplatin. Platinum preparations are excellent in terms of product development because they can easily detect accumulation in the fetus. Examples of the alkylating agent include nitrogen mustards such as cyclophosphamide, ifosfamide, melphalan, busulfan and thiotepa, and nitrosoureas such as nimustine, ranimustine, dacarbazine, procarbazine, temozolomide, carmustine, streptozotocin and bendamstin. Antimetabolites include fluorouracil, 6-mercaptopurine, azathioprine, hydroxyurea, thioguanine, fludarabine, cladribine, silatabin, gemcitabine and the like. Examples of topoisomerase inhibitors include camptothecin, irinotecan, nogitecan, anthracycline, doxorubicin, epirubicin, daunorubicin, bleomycin, levofloxacin, cyprofloxacin and the like. Examples of the microtubule polymerization inhibitor include vincristine, vindesine, paclitaxel, docetaxel and the like. Examples of endocrine therapeutic agents include anastrozole, exemestane, tamoxifen, toremifene, fulvestrant, letrozole and the like. In particular, from the viewpoint of being used as a therapeutic agent for breast cancer that frequently occurs in pregnant women, an endocrine therapeutic agent and the like can be mentioned. From the viewpoint of increasing safety, doxorubicin, cyclophosphamide, fluorouracil, etc., which have been proven to have less effect on the fetus, may be used.

In a specific aspect of the present invention, the present invention relates to a pharmaceutical composition for administration to a pregnant woman or a woman of childbearing potential, which comprises a dahaplatin derivative micelle coordinated to a polyethylene glycol polyamino acid block copolymer. In this dahaplatin derivative micelle, dahaplatin, which is a partial structure in which the oxalate group of oxaliplatin is removed, is coordinated to the carboxylate group of a polyamino acid (particularly glutamic acid or aspartic acid) instead of the oxalate group, and the micelle size is large. It is characterized in that it is 10 to 100 nm, preferably 20 to 90 nm, and more preferably 25 to 75 nm. Pharmaceutical compositions containing such dahaplatin derivative micelles may be administered in contraindicated doses and dosages to pregnant or potentially pregnant women when the active agent dahaplatin is administered. As an example, dahaplatin derivative micelles can be administered at 90-120 mg / m ² once weekly to pregnant or potentially pregnant women.

Anti-inflammatory drugs are non-steroidal anti-inflammatory drugs such as indomethacin, ketoprofen, loxoprofen, diclofenac, ibuprofen, acetylsalicylic acid, selecoxib, etodolac, and meloxicam. The pharmaceutical composition of the present invention containing an anti-inflammatory agent can be used as an analgesic and antipyretic drug, a therapeutic agent for rheumatoid arthritis, and can be used for the treatment or prevention of premature birth or miscarriage. Indomethacin and the like can be designed to be released in response to low pH in the inflamed area, as it is desired to act in the inflamed area. Further, the drug may be designed to be released by the action of esterase possessed by the macrophage by being phagocytosed by the macrophage in the inflamed area. In addition, the therapeutic agents for diseases that lead to inflammation such as endometriosis, the therapeutic agents for inflammation associated with infectious diseases, and the therapeutic agents for inflammation associated with immunity are also included in the anti-inflammatory agents in this document. be able to.

Examples of antihypertensive drugs include Ca antagonists, ARBs, ACE inhibitors, diuretics, β-blockers, α-blockers, and statin-based hyperlipidemia drugs. Calcium channel blockers include dihydropyridines such as amlodipine, nifedipine, nicardipine, benidipine, balnidipine, nitrendipine, nisoldipine, azelnidipine, manidipine, ephonidipine, silnidipine and alanidipine, benzothiazepines such as diltiazem, and phenyl such as verapamil. Examples include alkylamine-based agents. Examples of ARB include valsartan, losartan, candesartan, irbesartan, azilsartan and the like. Examples of the ACE inhibitor include imidapril, enalapril, delapril, silazapril, quinapril, temocapril, perindopril, lisinopril, trandolapril and the like. Examples of diuretics include trichloromethiazide, hydrochlorothiazide, furosemide, torasemide, azosemide, spironolactone, triamterene, eplerenone and the like. Examples of β-blockers include atenololol, bisoprolol, betaxolol, metoprolol, propranolol, nadolol, nipradilol, carteolol, bindrol, amosularol, arotinolol, carvedilol, labetalol, and bevantolol. Examples of the α blocker include uravidyl, terazosin, prazosin, doxazosin, and bunazosin. Examples of statins include simvastatin, pravastatin, lovastatin, pitavastatin, and atorvastatin. The pharmaceutical composition of the present invention including a statin drug can be used as an antihypertensive drug, an antihyperlipidemic drug, a therapeutic or prophylactic drug for preeclampsia. Since statins mainly inhibit cholesterol synthesis in the liver, it is preferable to design them to act in the liver.

In another aspect of the invention, the invention relates to a drug bound to a polyethylene glycol / polyamino acid block copolymer and micelles formed from the block copolymer. Here, the polyethylene glycol / polyamino acid block copolymer is a polyethylene glycol / polyamino acid block copolymer of the formula (I) or (II) defined in the present specification. As the drug bound to the polyethylene glycol / polyamino acid block copolymer, any of the above-mentioned drugs may be used, in particular, indomethacin and simvastatin.

When the therapeutic agent is indomethacin, indomethacin is bound via linker L ₃ in R ^2b and R ^2d of the block copolymers described above, and the carboxyl group of indomethacin is bound to linker L ₃ . As an example, R ^2b and R ^2d are aspartic acid side chains or glutamine side chains, and the chemical formulas in that case are shown below:

(In the formula, R ^1a , R ^1b , R ³ , R ⁴ , m, n, b are as defined herein.
c is 1 or 2 and L ₃ represents C _1- C ₆ alkylene)
The indomethacin-bound block copolymer according to the present invention can self-assemble in water to form micelles.

From the perspective of forming crosslinks within the micelles, furan may be introduced into some drug binding moieties for crosslinking. The furfuryl group in the crosslinked portion can be further crosslinked in the formation of micelles by further acting on a crosslinking agent such as bismaleimide. The compound of the present invention in which the therapeutic agent is indomethacin and a furfuryl group is introduced has the following structure as an example:

(During the ceremony
R ^1a , R ³ , R ⁴ , m, n, b are as already defined herein.
k represents the number of bridges introduced, k is an integer from 0 to 199,
c is 1 or 2 and L ₃ represents C _1- C ₆ alkylene. ). In another aspect of the present invention, the compound of the present invention having a flufuryl group introduced therein is made into micelles and crosslinked by the addition of 1,1- (methylenedi-4,1-phenylene) bismaleimide (BMI). Good.

If the therapeutic agent is simvastatin, simvastatin can be attached directly to the carboxy group of the side chain of the amino acid by an ester bond. Simvastatin-binding block copolymers are, for example:

(During the ceremony
R ^1a , R ³ , R ⁴ , m, n, b are as already defined herein.
c is 1 or 2).
Another aspect of the present invention relates to micelles formed by self-assembling the simvastatin-binding block copolymer according to the present invention in water.

The present invention also relates to a method for producing micelles containing a polyethylene glycol-polyamino acid block copolymer to which a therapeutic agent is bound or coordinated. The micelles are predominantly composed of polyethylene glycol-polyamino acid block copolymers to which the therapeutic agent is bound or coordinated, but may contain any other component as long as the desired size can be achieved. Good. Specifically, the method for producing micelles of the present invention is as follows:
A step of reconstitution of a polyethylene glycol-polyamino acid block copolymer to which a therapeutic agent is bound or coordinated in water, and
A step of obtaining micelles having a particle size of 20 to 100 nm by performing ultrafiltration using a fractionated molecular weight of 10,000 to 100,000 dialysis membranes.
including. Further, a step of purifying through a filter having a membrane size of 0.15 to 0.3 may be included. The size of micelles can be measured by using the dynamic scattering method.

All references referred to herein are incorporated herein by reference in their entirety.
The examples of the present invention described below are for illustration purposes only and do not limit the technical scope of the present invention. The technical scope of the present invention is limited only by the description of the claims. Changes to the present invention, for example, addition, deletion and replacement of the constituent elements of the present invention can be made provided that the gist of the present invention is not deviated.

Example 1: Evaluation of accumulation in placenta and fetus using polymer micelles of different sizes In this example, the accumulation of polymer micelles and low molecular weight compounds of different sizes in placenta and fetus was determined by using a pregnancy model mouse. It was evaluated by the fluorescence analysis, element analysis and mass spectrometry used.

Preparation of Dahaplatin-encapsulating micelles Two types of fluorescent-labeled dahaplatin-encapsulating micelles (Alexa-555 labeled micelles having a particle size of 30 nm and 70 nm and 30 nm particle size and Alexa647-labeled micelles having a particle size of 70 nm) were prepared (Non-Patent Document 2). : H. Cabral et. Al., Nat. Nanotech. 6 (2011) 815-823).

Administration to Pregnancy Model Mice A mixed solution of two types of dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm) was administered to mice 14 days after pregnancy. 48 hours after administration, the placenta and fetus were removed and frozen samples were prepared. Then, a frozen section (thickness: 10 mm) was prepared and nuclear-stained with Hoechst. Fluorescence from Hoechst (FIG. 1A) and fluorescence from micelle labels (Alexa 555 and Alexa 647) (FIGS. 1B and C) were measured. Next, the distribution of platinum, iron, potassium, and calcium in frozen sections of the fetus obtained from pregnant mice administered with dahaplatin-encapsulated micelles (particle size 30 nm) was measured by a synchrotron (Spring-8) (Fig. 2). .. From the results shown in FIG. 1, it was confirmed that two types of dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm) were not accumulated in the fetus (F). In the dahaplatin-encapsulated micelles having a particle size of 30 nm, accumulation in the fetus (F) was not observed, but accumulation of micelles was observed up to the labyrinth (L) layer on the fetal side of the placenta. In dahaplatin-encapsulated micelles with a particle size of 70 nm, accumulation up to the maternal dedidua (D) layer of the placenta was observed, but spongy trophoblast cells (spongiotrophoblast) between the D and L layers ( No accumulation in the S) layer was observed. From this result, it was shown that the distribution in the placenta can be controlled by precisely designing the size of the drug. From FIG. 2, since the platinum element derived from dahaplatin was not found in the fetus, it was shown that the passage of the placenta was suppressed by increasing the size of the drug to 30 nm or more.

On the 14th day after pregnancy, two types of dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm) and oxaliplatin (8 mg / kg), which is an active form of dahaplatin, were administered, respectively. 48 hours after administration, the placenta and fetus were removed, 1 mL of lysis buffer (Promega, USA) was added to the removed placenta and fetus, and after homogenization, platinum elements in the tissue were quantified by inductively coupled plasmon mass spectrometry (Fig. 3). : Fetus, Figure 4: Placenta). From the results shown in FIG. 3, in the fetuses of pregnant mice treated with oxaliplatin, the accumulation of platinum in the fetuses of platinum was significantly higher than that of the fetuses of pregnant mice treated with dahaplatin-encapsulated micelles, and the particle size was 30 nm. It was shown that the above results suppressed the passage of particles through the placenta. From the results shown in FIG. 4, platinum was accumulated in the placenta between the fetuses of pregnant mice treated with oxaliplatin and the fetuses of pregnant mice treated with dahaplatin-encapsulated micelles (particle size 30 nm and 70 nm). Compared with the oxaliplatin administration group, the accumulation was decreased in the administration group of micelle having a particle size of 30 nm, and further decreased in the administration group of micelle having a particle size of 70 nm. From this, it was shown that the placental accumulation can be controlled by precisely designing the size of the drug.

Example 2-1: Evaluation of permeability to human placenta In this example, the permeability of polymer micelles, low molecular weight compounds, and polymers of different sizes in human placenta was evaluated using a human placenta perfusion model.

Preparation of Drugs According to the preparation of dahaplatin-encapsulating micelles of Example 1, fluorescent-labeled dahaplatin-encapsulating micelles having particle sizes of 30 nm and 70 nm were prepared. 8-branched PEG (molecular weight: 40 kDa, 10 nm size (dynamic light scattering method), 20 mg, 0.50 μmol) having an azide group at the terminal was dissolved in DMSO (1.0 mL, 20 mg / mL) and Cy3-DBCO (7). (0.0 mg, 6.0 μmol) was added and reacted at −20 ° C. overnight. Then, dialysis (MWCO: 6,000-8,000) with pure water and freeze-drying gave Cy3-labeled 8armPEG.

Regarding the human placental perfusion model, Non-Patent Document 3 (K. Shintaku et. Al., Drug Metab. Dispos. 37 (2009) 962) Non-Patent Document 4 (J. R. Huston et. Al., Clin. Pharmacol. Ther . 90 (2011) 67). Specifically, a needle (18 gauge) was introduced into the maternal side of the human placenta provided by the Department of Obstetrics and Gynecology at the University of Tokyo Hospital, and a needle (18 gauge) was introduced into the vein and artery on the fetal side, respectively. Krebs-Ringer carbonate buffer prepared according to M. Nagai et. Al., Drug Metab. Dispos. 41 (2013) 2124) was perfused at 37 ° C for 30 minutes (maternal flow velocity: 15 mL / min, fetal flow velocity). : 3 ml / min). Here, the buffer was saturated with oxygen generated by the redox reaction of hydrogen peroxide solution and iron chloride. Then, dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm) (dahaplatin dose: 85.4 mg / L), oxaliplatin (2.7 mg / L) and Cy3-labeled 8 armPEG (fluorescence intensity: 3,000 (RFU)). Perfusion was performed for 60 minutes, and 1 mL of the solution from the vein on the fetal side was collected every 5 minutes. For the oxaliplatin and dahaplatin-encapsulated micelle-administered groups, the platinum content in the collected samples was measured by inductively coupled plasmon mass spectrometry. For the polymer, the fluorescence intensity was quantified by the HPLC method. The ratio of the permeation amount to the dose of each drug was calculated and shown in FIG. For the oxaliplatin and dahaplatin-encapsulating micelle-administered groups, after the experiment, 2 mL of 90% nitric acid aqueous solution was added to the placenta, dissolved in 2 mL of 1% nitric acid aqueous solution, and the platinum content in the placenta was quantified by inductively coupled plasmon mass spectrometry. did. The platinum content obtained was standardized by the mass of the placenta (Fig. 6). From the results shown in FIG. 5, oxaliplatin permeated the placenta, but neither dahaplatin-encapsulating micelles (particle size 30 nm and 70 nm) permeated the placenta. Therefore, by increasing the drug size to 30 nm or more, the placenta permeated. It was suggested that it was suppressed. In addition, placental permeation was observed in 8-armPEG, although it was lower than that in oxaliplatin, suggesting that there was a threshold between 10 nm and 30 nm for drug size. From the results of FIG. 6, it was shown that oxaliplatin accumulated in the placenta and micelles did not accumulate in the placenta, and thus the accumulation of micelles in the placenta was suppressed due to the difference in placenta structure between mice and humans. As a result, it was confirmed that the accumulation and permeation into the placenta can be suppressed by increasing the size of the drug to 30 nm or more.

Example 2-2: Assessment of permeability to human placenta To identify the size of material that passes through the placenta, PEG-coated gold nanoparticles (NPs) at 10, 20, and 30 nm (5 mL, Nanocs, Inc.). , USA), an ex vivo human placenta perfusion experiment was performed in the same manner as in Example 2-1. When 10 nm gold NP was perfused, the amount of gold nanoparticles detected from the fetal side increased in a time-dependent manner, reaching 4.58% of the dose after 60 minutes. In addition, when 20 nm Gold NP was perfused, the detected dose from the fetal side was 2.21%. On the other hand, when 30 nm Gold nanoparticles were perfused, the detected dose from the fetal side was 0.05% of the administered dose after 60 minutes. The results are shown in FIG. 5B. These results suggest that there may be a size cutoff between 20 nm and 30 nm for the material to pass through the placenta.

Example 3: Preparation of indomethacin-encapsulating micelles for the treatment of preterm birth In this example, block copolymers were synthesized and micelles were prepared for the construction of indomethacin-encapsulating micelles.
Experimental Method According to a previously reported report (Non-Patent Document 6: M. Yokoyama et. Al., Makromol. Chem. 190 (1989) 2041-2054), an NCA polymerization method using PEG (molecular weight: 12,000 Da) primary amine as an initiator. PEG-poly (β-benzyl-L-apartate) (PEG-PVLA) was synthesized. Then, according to the following scheme, PEG-poly (Asp-Furan-OH) (PEG-P (Asp-furan-OH)) is produced by an aminolysis reaction of 4-amino-1-butanol and 2-aminomethylfuran with the BPLA chain. Synthesized.

Specifically, PEG-PVLA was dehydrated by the benzene freeze-drying method and dissolved in distilled DMF (100 mg / mL). Then, 2 equivalents of distilled 4-amino-1-butanol and 2-aminomethylfuran were added to the BPLA chain, and the mixture was reacted at room temperature for 6 hours. After the reaction, the reaction solution was added to an excess amount of diethyl ether to give a polymer. Next, indomethacin was introduced into the polymer by esterifying the hydroxyl group in the polymer with indomethacin. Specifically, dimethylaminopyridine (DMAP), water-soluble carbodiimide (WSC) and indomethacin were mixed (DMAP / WSC / indomethacin = 2: 3: 1 (mass ratio)) and dissolved in DMF (1 mg / mL). .. Then, the solution was added to PEG-P (Asp-furan-OH) so that the molar ratio of indomethacin to the BPLA chain was 4, and the reaction was carried out overnight at room temperature. Then, the reaction solution was added to an excess amount of diethyl ether to obtain a polymer. The composition of the obtained polymer was measured by ^{1 1} H-NMR. ^{From 1 1} H-NMR, it was confirmed that the introduction rate of 4-amino-1-butanol was 80% and the introduction rate of 2-aminomethylfuran was 20%.

Indomethacin-encapsulating micelles were prepared by dialysis. The polymer was dissolved in DMSO (2,7,14 mg / mL), dialyzed against pure water (MWCO: 3,500 Da), and purified by a filter (0.22 μm). Then, 1,1- (methylenedi-4,1-phenylene) bismaleimide (BMI) was added to the micelle solution, and the reaction was carried out at 50 ° C. for 2 days. After the reaction, it was purified by ultrafiltration (MWCO: 100,000 Da) and a filter (0.22 μm). The size and polydispersity (PDI) of the prepared micelles were measured by dynamic light scattering (DLS).

From FIG. 7, it was confirmed that the degree of polymerization of the PAsp chain was 32, the amount of indomethacin introduced was 16 (/ polymer), and the amount of furan introduced was 6 (/ polymer). According to DLS measurement, micelles prepared at polymer concentrations of 2, 7, and 14 mg / mL are 41 nm (PDI: 0.09), 37 nm (PDI: 0.04), and 39 nm (PDI: 0.04) sizes, respectively. Was confirmed, suggesting that the polymer concentration during micelle preparation does not significantly affect the size of micelles. Moreover, since the size after BMI cross-linking was 40 nm from Table 1, it was confirmed that the size did not change before and after the cross-linking.

Example 4: Stability test of indomethacin-encapsulating micelles In this example, as an index of the stability of indomethacin-encapsulating micelles, the scattered light intensity of the micelle after the addition of the surfactant, and the physiological salt condition and the pH condition in the endosome are used. Indomethacin release amount was measured.
Experimental method Sodium dodecylsulfonate (SDS), which is one of the surfactants, was added to the micelle solution as an index of stability. Specifically, SDS (20 g) was dissolved in pure water (80 mL) and stirred at 65 ° C. Then, pure water (100 mL) was added to the SDS solution to prepare a 20% SDS aqueous solution. The prepared 20% SDS aqueous solution (10 μL) was added to the micellar solution (70 μL), and after stirring, the size, polydispersity and scattered light intensity (divided count rate (DCR)) were measured by DLS.
In addition, the amount of indomethacin released under physiological salt conditions and endosome pH conditions was used as an index of stability. Specifically, indomethacin-encapsulating micelles were dialyzed against pure water (MWCO: 3,500 Da), and the amount of indomethacin in the sample after dialysis was measured by the HPLC method.

From Table 2, while the scattered light intensity of micelles (NCL) before BMI cross-linking decreased due to the addition of SDS, the scattered light intensity of micelles (CL) after cross-linking did not decrease even after the addition of SDS. It was suggested that the stability of micelles was improved. In addition, FIG. 8 showed no release of indomethacin at blood pH (pH 7.4) and endosome pH (pH 5.3), suggesting that indomethacin can be stably included in blood and endosomes. ..

Example 5: Indomethacin release test in macrophages In this example, the release of indomethacin from indomethacin-encapsulating micelles was evaluated by measuring the amount of indomethacin in macrophage cells.

Experimental method For culturing macrophage cells (RAW246.7 cells), cells were seeded at a concentration of 200,000 cells / mL in a 6-well plate for cell culture, and contained 10% fetal bovine serum (FBS) and 1% penicillin / streptomycin. The cells were cultured in DMEM for 24 hours. Then, indomethacin-encapsulating micelles prepared in Example 4 (indomethacin amount: 0.5 mg / mL: micelle diameter: about 40 nm, crosslinked) and indomethacin alone (0.5 mg / mL) were added, respectively, 24, 48, After 72 hours, the cells were washed with PBS and 2% SDS (1 mL) was added to prepare a cell suspension. The amount of indomethacin in the solution was measured by the HPLC method (Fig. 9). From FIG. 9, since the amount of indomethacin released from the indomethacin-encapsulating micelle (CL) is about the same as the amount of uptake of indomethacin alone (FD), the ester bond in the micelle is cleaved by the ester-degrading enzyme in macrophages, and as a result. Efficient release of indomethacin was suggested.

Example 6: Evaluation of permeability to human placenta using indomethacin-encapsulating polymer micelles In this example, the permeability of polymer micelles, low molecular weight compounds, and polymers of different sizes in human placenta was measured using a human placenta perfusion model. Evaluated.

Experimental method For the human placental perfusion model, Non-Patent Document 3 (K. Shintaku et. Al., Drug Metab. Dispos. 37 (2009) 962) Non-Patent Document 4 (JR Huston et. Al., Clin. Pharmacol. Ther. It was prepared according to 90 (2011) 67). Specifically, a needle (18 gauge) was introduced into the maternal side of the human placenta provided by the Department of Obstetrics and Gynecology at the University of Tokyo Hospital, and a needle (18 gauge) was introduced into the vein and artery on the fetal side, respectively. The Krebs-Ringer carbonate buffer prepared according to M. Nagai et. Al., Drug Metab. Dispos. 41 (2013) 2124) was perfused at 37 ° C for 30 minutes (maternal flow velocity: 15 mL / min, fetal flow velocity). : 3 ml / min). Here, the buffer was saturated with oxygen generated by the redox reaction of hydrogen peroxide solution and iron chloride. Then, the indomethacin-encapsulating micelle (indomethacin amount: 2 mg / L: micelle diameter: about 40 nm, crosslinked) prepared in Example 4 and indomethacin alone (2 mg / L) were perfused for 60 minutes, and the solution from the vein on the fetal side. Was collected every 5 minutes. The amount of indomethacin in the collected sample was quantified by the HPLC method (Fig. 10). When indomethacin (IND) was refluxed, the pass rate after 60 minutes was 17.6% (n = 3). On the other hand, when indomethacin-encapsulating micelles (IND / m) were refluxed, the passage rate after 60 minutes was 0.03% (n = 3) (p <0.05). From the results of FIG. 10, since indomethacin alone penetrated the placenta but micelles did not penetrate the placenta, it was suggested that the passage of the drug through the human placenta was suppressed by increasing the size of the drug to 30 nm or more. It was.

Example 6-2: Evaluation of tissue migration of indomethacin-encapsulating polymer micelles On the morning of the 18th and 19th days of pregnancy, fluorescently labeled indomethacin micelles (IND dose / kg, 10 mg) were administered from the tail vein of pregnant mice. After 1 hour, 4 hours, 8 hours, and 24 hours, mice are euthanized and the brain, lungs, heart, liver, spleen, pancreas, both kidneys, fetus, umbilical cord, placenta, amniotic membrane, cervix, The cervix, fetus muscle, and femur were harvested and drug distribution was evaluated using the IVIS imaging system. FIG. 11A shows the location of the recovered organs. 11B-E show fluorescence at 1 hour, 4 hours, 8 hours, and 24 hours after administration. These experiments showed that indomethacin micelles were distributed in the liver, kidneys, spleen, uterine body, and placenta, and did not reach the fetus at each point in time.

Example 6-3: Evaluation of tissue migration of indomethacin-encapsulating polymer micelles Indomethacin (IND dose / kg as 1 mg) and was administered from the tail vein of pregnant mice on the morning of the 18th and 19th days of pregnancy. After 1 hour, 4 hours, 8 hours, and 24 hours, the mice were euthanized and the fetus, placenta, uterine body, uterus, kidneys, liver and spleen were collected. Samples were homogenized and indomethacin concentrations in tissues were measured by HPLC. In the indomethacin micelle-administered group, free indomethacin in tissues and indomethacin in micelle state were detected as the total amount. This is because indomethacin micelles decompose when sodium hydroxide is used for homogenization. In the free indomethacin group, 0.57% of the maternal dose was detected in the fetus after 24 hours, which was the maximum. On the other hand, in the indomethacin micelle group, only 0.013% of the maternal dose was distributed to the fetus, which was also the maximum after 24 hours. Therefore, administration of indomethacin micelles significantly reduced (E) distribution to the fetus compared to administration of free indomethacin (p <0.05), thereby preventing indomethacin micelles from crossing the mouse placenta. It has been shown. Furthermore, in organs such as (A) kidney, (B) liver, (C) spleen, and (D) placenta, the amount of indomethacin detected within 24 hours was not sufficient to cause organ damage (Fig. 12A-E).

Example 7: Therapeutic effect of indomethacin-encapsulating micelles using preterm birth model mice In this example, the preterm birth inhibitory effect and toxicity of indomethacin-encapsulating micelles were evaluated using preterm birth model mice.

Experimental method Preterm model mice were prepared by administering lipopolysaccharide (LPS) to the cervix according to the previous report (L. L. Reznikov et. Al., Biol. Reprod. 60 (1999) 1231-1238). Specifically, on the 15th day of pregnancy, LPS (3 μg / mouse) was administered to the vaginal lumen via a soft tube for oral administration of mice.
Then, on days 15, 16, 17 and 18 of pregnancy, indomethacin-encapsulating micelles (10 mg / kg: micelle diameter: about 40 nm, with / without cross-linking) and indomethacin alone (1 mg / kg) were administered until the 19th day of pregnancy. Delivery was premature. In addition, the abdomen was opened on the 19th day of pregnancy, and the survival rate of the fetus was measured. The results are shown in FIG.

Table 3 showed that administration of micelles and indomethacin alone reduced the number of preterm mice, suggesting that indomethacin suppresses inflammation in the cervix. In addition, the administration of micelles increased the survival rate of the fetus, suggesting that the increase in drug size decreased the amount of accumulation in the fetus.

Example 8: Preparation of simvastatin-encapsulating micelles for the treatment of preeclampsia In this example, simvastatin was introduced into a block copolymer and micelles were prepared for the treatment of preeclampsia.

Experimental method According to Non-Patent Document 7 (A. Harada and K. Kataoka, Macromolecules 28 (1995) 5294-5299), PEG-poly (β) was produced by an NCA polymerization method using a primary amine of PEG (molecular weight: 12,000 Da) as an initiator. -Benzyl-L-aspartate) (PEG-PVLA) was synthesized, and PEG-poly (L-aspartate) (PEG-PAsp) was synthesized by deprotecting the benzyl group of the side chain structure. Simvastatin was introduced into the polymer by esterifying the carboxyl group of PEG-PAsp and the hydroxyl group of simvastatin according to the scheme below.

PEG-PAsp was dissolved in DMF (10 mg / mL), 10 equivalents of DMAP, EDC and simvastatin were added to the carboxyl group of the PAsp chain, and the reaction was carried out at room temperature for 24 hours. Then, the reaction solution was added to an excess amount of diethyl ether to obtain PEG-P (Asp-simvastatin). ^{1 From 1} H-NMR, the amount of simvastatin introduced was 1.2 mg / (mg polymer).

Simvastatin-encapsulating micelles were prepared by the dialysis method. The polymer was dissolved in DMAc (1 mg / mL), dialyzed against pure water (MWCO: 3,500 Da), and purified by a filter (0.22 μm). The size and polydispersity (PDI) of the prepared micelles were measured by dynamic light scattering (DLS).

Result ^{1 From 1} H-NMR, the amount of simvastatin introduced was 1.2 mg / (mg polymer). Further, it was confirmed by DLS measurement that micelles having a size of 35 nm and a PDI of 0.23 were obtained.

Example 9: Therapeutic effect and toxicity test of simvastatin-encapsulating micelles using preeclampsia model mice In this example, the antihypertensive effect and toxicity of simvastatin-encapsulating micelles were evaluated using preeclampsia model mice.

Experimental method Preeclampsia model mice were prepared by the method of administering angiotensin II (AngII) according to the previously reported (C.C. Zhou et. Al., Nat. Med. 14 (2008) 855-862). Specifically, from the 10th to the 17th day of pregnancy, AngII (1.5 g / kg) was administered to pregnant mice via a subcutaneous pump (flow velocity: 1.0 μL / h) to prepare a model of hypertension.

Simvastatin-encapsulating micelles (20 μg / kg: Simvastain Micelle), simvastatin alone (20 μg / kg: Simvastatin), and pravastatin (20 μg / kg: Pravastatin) were administered on the 10th to 17th days of pregnancy, and the drug-free group (NS) The maternal blood pressure, urinary protein content and fetal weight were evaluated in comparison with. Blood pressure was evaluated by a sphygmomanometer on days 10, 13, 15, and 17 of pregnancy (Fig. 14). Regarding the body weight of the fetus (FBW (g)), the fetus was removed on the 17th day of pregnancy and the body weight was measured (FIG. 15). For urinary protein, the amount of albumin in urine on days 10, 11, 16 and 17 of pregnancy was measured (Fig. 16).

Results From FIG. 14, since the blood pressure in the drug-administered group was lower than that in the non-administered group, it was confirmed that the statin preparation reduced the blood pressure. In addition, the blood pressure in the simvastatin-encapsulated micelle group was similar to that before the administration of Ang II, suggesting the effect of cleavage of the ester bond in the micelle and the resulting sustained release of the drug. From FIG. 15, the weight of the fetus in the simvastatin-encapsulating micelle group (AngiI-Simvastatin Micelle) was similar to that in the Ang II-non-administered group (NS-NS), suggesting that the placental function was improved by eliminating hypertension. It was also suggested that the size increased due to micelle formation, and as a result, the amount accumulated in the fetus decreased. From FIG. 16, the increase in urinary albumin level was suppressed in the simvastatin-encapsulating micelle group, suggesting improvement of renal function by eliminating hypertension.

Claims (21)

1. A pharmaceutical composition for administration to a pregnant woman or a woman of childbearing potential, which comprises particles having a hydrophilic polymer on the surface to which a therapeutic agent is bound or coordinated, and has a particle size of a dynamic light scattering method. The pharmaceutical composition having a diameter of 10 to 100 nm as measured by.
2. The pharmaceutical composition according to claim 1, wherein the particles are micelles containing a polyethylene glycol-polyamino acid block copolymer.
3. The polyethylene glycol-polyamino acid block copolymer has the following general formula (I) or (II):
   

   

   (During the ceremony
   R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
   L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
   L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
   R ^2a and R ^2c , respectively, independently exist at each appearance or represent a methylene group.
   R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
   R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
   R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
   m is an integer from 20 to 2,000
   n is an integer from 1 to 200)
   2. The block copolymer represented by, wherein the therapeutic agent is coordinated via R ^2b and R ^2d of the block copolymer, or is bound directly or via a linker. The pharmaceutical composition according to.
4. The pharmaceutical composition according to any one of claims 1 to 3, wherein the particle size is 25 to 75 nm.
5. The pharmaceutical composition according to any one of claims 1 to 4, wherein the therapeutic agent is administered in a dosage and / or dosage contraindicated for a pregnant woman when administered only with the therapeutic agent.
6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the therapeutic agent is selected from the group consisting of an anticancer drug, an anti-inflammatory drug, an antihypertensive drug, and a psychotropic drug.
7. The pharmaceutical composition according to claim 6, wherein the anticancer drug is a platinum preparation.
8. The pharmaceutical composition according to claim 7, wherein the platinum preparation is formed through a coordination bond with one or two carboxylate groups of the amino acid side chain.
9. The pharmaceutical composition according to any one of claims 1 to 6, wherein the therapeutic agent is an anti-inflammatory agent and is for preventing miscarriage.
10. The pharmaceutical composition according to claim 9, wherein the anti-inflammatory drug is a non-steroidal anti-inflammatory drug.
11. The pharmaceutical composition according to any one of claims 1 to 6, wherein the therapeutic agent is an antihypertensive agent and is for the treatment of a gestational hypertension agent.
12. The pharmaceutical composition according to claim 11, wherein the antihypertensive drug is a statin-based antihypertensive drug.
13. The pharmaceutical composition according to any one of claims 1 to 12, wherein the pharmaceutical composition is a pharmaceutical composition for reducing fetal phytotoxicity.
14. The pharmaceutical composition according to any one of claims 1 to 12, wherein the pharmaceutical composition is a pharmaceutical composition for reducing placental barrier permeation.
15. The following general formula (I) or (II):
    

    

    (During the ceremony
    R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
    L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
    L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
    R ^2a and R ^2c , respectively, independently exist at each appearance or represent a methylene group.
    R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
    R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
    R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
    m is an integer from 20 to 2,000
    n is an integer from 1 to 200)
    A drug is a drug-combined block copolymer in which a drug is bound to the block copolymer represented by (1) via R ^2b and R ^2d of the block copolymer, directly or via a linker, and the drug is , Indomethacin or simvastatin, said drug complex block copolymer.
16. 15. The pharmaceutical composition comprising micelles formed from the sound drug complex block copolymer according to claim 15, wherein the particle size is 20 to 100 nm when measured by a dynamic light scattering method. object.
17. The pharmaceutical composition according to claim 16, which contains indomethacin as the drug and the pharmaceutical composition is a miscarriage-preventing pharmaceutical composition.
18. The pharmaceutical composition according to claim 16, which contains simvastatin as the drug and the pharmaceutical composition is a pharmaceutical composition for the treatment of preeclampsia.
19. A method for producing micelles containing a polyethylene glycol-polyamino acid block copolymer to which a therapeutic agent is bound or coordinated and for administration to a pregnant woman or a woman of childbearing potential.
    A step of reconstitution of the polyethylene glycol-polyamino acid block copolymer in water, and
    A step of obtaining micelles having a particle size of 20 to 100 nm by performing ultrafiltration using a dialysis membrane having a molecular weight cut off of 10,000 to 100,000.
    The production method.
20. The production method according to claim 19, wherein the therapeutic agent is selected from the group consisting of an anticancer drug, an anti-inflammatory drug, an antihypertensive drug, and a psychotropic drug.
21. The polyethylene glycol-polyamino acid block copolymer has the following general formula (I) or (II):
    

    

    (During the ceremony
    R ^1a and R ^1b represent hydrogen atoms, hydroxyl groups or unsubstituted or substituted linear or branched C _1-12 alkyl groups or C _1-12 alkoxy groups.
    L ¹ represents-(CH ₂ ) _b- NH-, and b is an integer of 1 to 5.
    L ² represents − (CH ₂ ) _c −CO−, and c is an integer from 1 to 5.
    R ^2a and R ^2c , respectively, independently exist at each appearance or represent a methylene group.
    R ^2b and R ^2d represent a carboxy group or any amino acid side chain.
    R ³ represents a hydrogen atom, protecting group, hydrophobic group or polymerizable group.
    R ⁴ represents a hydroxyl group, an oxybenzyl group, an -NH- (CH ₂ ) _a- X group or an initiator residue, where a is an integer of 1-5 and X is a primary, secondary, An amine compound residue containing one or more of the tertiary amine or quaternary ammonium salt, or a compound residue that is not an amine.
    m is an integer from 20 to 2,000
    n is an integer from 1 to 200)
    A block copolymer represented by, wherein the therapeutic agent is bound to the R ^2b and R ^2d groups of the block copolymer, or is bound directly or via a link via a linker. Item 19. The production method according to Item 19.

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