JP7244821B2 - Novel compound having bone metabolism inhibitory activity and method for producing the same - Google Patents

Description

TECHNICAL FIELD The present invention relates to a novel compound having bone metabolism inhibitory activity and a method for producing the same.

Bone is a dynamic tissue that maintains a balanced state of bone metabolism through constant repetition of bone formation by osteoblasts and bone resorption by osteoclasts. It is known that when the functional balance of osteoblasts and osteoclasts is abnormal, the dynamic balance of bone metabolism is disrupted, causing various diseases caused by abnormal bone metabolism.

Factors that control bone formation include bone morphogenetic proteins (BMPs). BMPs are cytokines that induce bone. BMP signals are transduced into cells by type I and type II serine-threonine kinase receptors, and further transduced into the nucleus by phosphorylation of the transcriptional regulator Smad by type I receptors. It has become clear that a lack of BMP signals causes brachydactyly or achondroplasia, while an excess of BMP signals causes fibrodysplasia ossificans progressive and the like (Non-Patent Document 1). . In addition, BMPs play essential roles in various physiological bone formations such as skeletal formation and fracture healing. Therefore, the range of clinical applications of BMP is wide and highly useful, including bone remodeling, treatment of jaw deformity, diagnosis of osteogenic diseases, and the like, and has attracted attention in recent years.

Fibrodysplasia ossificans progressive (FOP) is a disease in which tissue such as muscle, tendon or ligament turns into bone. Hallux valgus is common in patients from birth, but no other remarkable abnormalities are observed at birth. The first symptoms leading to osseous formation of FOP often occur by the age of 10 years. Its symptoms are swelling or hardening under the skin, ectopic ossification while repeating a phenomenon called flare-up, sometimes accompanied by heat or pain, resulting in worsening movement of the joints of the limbs or deformity of the back. In addition, it is impossible to surgically remove the resulting bone tissue because flare-ups are triggered by injury, surgery, or the like. Patients generally become incapacitated by the age of 30 and rarely live past the age of 40. The number of patients is said to be 1 in 2 million people, but the exact number is unknown. Since the number of patients is not so large, the onset mechanism has hardly been elucidated, and no treatment has been established (Non-Patent Document 2).

Recently, in patients with FOP, activin receptor-like kinase-2 (ALK2), a BMP type I receptor, has been constitutively activated by a point mutation of 206 arginine to histidine. was discovered. This indicated that the ALK2 receptor-mediated BMP signaling pathway is the main cause of FOP (Non-Patent Document 1).

In view of such findings, low-molecular-weight compounds that inhibit bone differentiation by inhibiting BMP signaling were searched, and dorsomorphin was found as a compound having the desired activity (Non-Patent Document 3). Furthermore, various derivatives were synthesized based on dorsomorphin, and a more excellent group of compounds was discovered (Non-Patent Document 4). Oral administration of LDN-193189, a kind of synthetic derivative of dorsomorphin, to FOP pathological model mice has been shown to significantly suppress the onset of FOP (Non-Patent Document 5).

Screening using iPS cells generated from FOP patient-derived cells showed that rapamycin can inhibit ectopic ossification (Non-Patent Document 6).

In addition, as a compound that inhibits osteogenesis, FKI-5513 substance was found from a fungal culture medium of the genus Trichoderma (Patent Document 1).

Patent document 2 describes a compound represented by formula (P-I), a salt thereof, or a solvate thereof as a novel compound having an inhibitory effect on BMP signaling via the ALK2 receptor.

Patent No. 5725469 Japanese Patent Application Laid-Open No. 2017-119691

Kaplan et al., J. Bone Miner Metab. 2008, vol.26, p.521-530 Shore et al., Nat. Genet. 2006, vol.38, p.525-527 Yu et al., Nat. Chem. Biol. 2008, Vol. 4, p. 33-41 Cuny et al., Bioorg. Med. Chem. Lett. 2008, 18, pp. 4388-4392 Yu et al., Nat. Med. 2008, vol. 14, p. 1363-1369 Hino et al., J. Clin. Invest. 2017, Vol. 127(9), p. 3339-3352

The ALK2 receptor-mediated BMP signaling pathway is thought to be not only a major cause of FOP, but also of relevance in other disorders of bone metabolism. Therefore, compounds that inhibit BMP signaling are believed to be useful in the prevention or treatment of many diseases caused by abnormal bone metabolism, including FOP. Compounds capable of inhibiting bone metabolism through their inhibitory activity on BMP signaling are known, but there is a need for more useful compounds from the viewpoint of pharmacological activity and/or safety.

Therefore, an object of the present invention is to provide novel compounds having bone metabolism inhibitory activity, which are highly pharmacologically active and safe.

The present inventors have studied various means for solving the above problems. The present inventors have found that a novel microorganism produces in its culture medium a compound having inhibitory activity on BMP signaling. The present inventors have found that the compound in the culture solution is a novel compound having a skeletal structure different from that of known bone metabolism inhibitors. In addition, the present inventors have found that the compounds can be produced by culture means using the microorganisms. The present inventors completed the present invention based on the above findings.

［式中、
R¹は、水素、ハロゲン（フッ素、塩素、臭素又はヨウ素）、シアノ、ニトロ、ヒドロキシル、置換若しくは非置換のアルキル、置換若しくは非置換のアルケニル、置換若しくは非置換のアルキニル、置換若しくは非置換のシクロアルキル、置換若しくは非置換のシクロアルケニル、置換若しくは非置換のシクロアルキニル、置換若しくは非置換のヘテロシクロアルキル、置換若しくは非置換のシクロアルキルアルキル、置換若しくは非置換のヘテロシクロアルキルアルキル、置換若しくは非置換のアリール、置換若しくは非置換のアリールアルキル、置換若しくは非置換のヘテロアリール、置換若しくは非置換のヘテロアリールアルキル、置換若しくは非置換のアルコキシ、置換若しくは非置換のシクロアルコキシ、置換若しくは非置換のヘテロシクロアルコキシ、置換若しくは非置換のアリールオキシ、置換若しくは非置換のアリールアルキルオキシ、置換若しくは非置換のアリールアルケニルオキシ、置換若しくは非置換のヘテロアリールオキシ、置換若しくは非置換のヘテロアリールアルキルオキシ、置換若しくは非置換のアルコキシカルボニル、置換若しくは非置換のシクロアルコキシカルボニル、置換若しくは非置換のアシル、置換若しくは非置換のアシルオキシ、又は置換若しくは非置換のアミノであり、
R²及びR³は、互いに独立して、水素、ハロゲン（フッ素、塩素、臭素又はヨウ素）、シアノ、ニトロ、ヒドロキシル、置換若しくは非置換のアルキル、置換若しくは非置換のアルケニル、置換若しくは非置換のアルキニル、置換若しくは非置換のシクロアルキル、置換若しくは非置換のシクロアルケニル、置換若しくは非置換のシクロアルキニル、置換若しくは非置換のヘテロシクロアルキル、置換若しくは非置換のシクロアルキルアルキル、置換若しくは非置換のヘテロシクロアルキルアルキル、置換若しくは非置換のアリール、置換若しくは非置換のアリールアルキル、置換若しくは非置換のヘテロアリール、置換若しくは非置換のヘテロアリールアルキル、置換若しくは非置換のアルコキシ、置換若しくは非置換のシクロアルコキシ、置換若しくは非置換のヘテロシクロアルコキシ、置換若しくは非置換のアリールオキシ、置換若しくは非置換のアリールアルキルオキシ、置換若しくは非置換のアリールアルケニルオキシ、置換若しくは非置換のヘテロアリールオキシ、置換若しくは非置換のヘテロアリールアルキルオキシ、置換若しくは非置換のアルコキシカルボニル、置換若しくは非置換のシクロアルコキシカルボニル、置換若しくは非置換のアシル、置換若しくは非置換のアシルオキシ、又は置換若しくは非置換のアミノであり、
R⁴は、水素、置換若しくは非置換のアルキル、置換若しくは非置換のアルケニル、置換若しくは非置換のアルキニル、置換若しくは非置換のシクロアルキル、置換若しくは非置換のシクロアルケニル、置換若しくは非置換のシクロアルキニル、置換若しくは非置換のヘテロシクロアルキル、置換若しくは非置換のシクロアルキルアルキル、置換若しくは非置換のヘテロシクロアルキルアルキル、置換若しくは非置換のアリール、置換若しくは非置換のアリールアルキル、置換若しくは非置換のヘテロアリール、置換若しくは非置換のヘテロアリールアルキル、置換若しくは非置換のアルコキシカルボニル、置換若しくは非置換のシクロアルコキシカルボニル、又は置換若しくは非置換のアシルである。］
で表される化合物若しくはその塩、又はそれらの溶媒和物。
（2） R¹が、ヒドロキシル、置換若しくは非置換のC₁～C₆アルコキシ、置換若しくは非置換のC₃～C₆シクロアルコキシ、置換若しくは非置換の3～6員ヘテロシクロアルコキシ、置換若しくは非置換のC₆～C₁₈アリールオキシ、置換若しくは非置換のC₇～C₂₀アリールアルキルオキシ、置換若しくは非置換のC₇～C₂₀アリールアルケニルオキシ、置換若しくは非置換の5～15員のヘテロアリールオキシ、置換若しくは非置換の5～15員のヘテロアリール-C₁～C₆アルキルオキシ、又は置換若しくは非置換のC₁～C₂₀アシルオキシである、前記実施形態（1）に記載の化合物若しくはその塩、又はそれらの溶媒和物。
（3） R²が、水素であり、且つR³が、置換若しくは非置換のC₁～C₆アルキル、置換若しくは非置換のC₂～C₆アルケニル、置換若しくは非置換のC₂～C₆アルキニル、置換若しくは非置換のC₃～C₆シクロアルキル、置換若しくは非置換のC₄～C₆シクロアルケニル、置換若しくは非置換のC₄～C₆シクロアルキニル、置換若しくは非置換の3～6員のヘテロシクロアルキル、置換若しくは非置換のC₇～C₁₁シクロアルキルアルキル、置換若しくは非置換の3～6員のヘテロシクロアルキル-C₁～C₆アルキル、置換若しくは非置換のC₆～C₁₈アリール、置換若しくは非置換のC₇～C₂₀アリールアルキル、置換若しくは非置換の5～15員のヘテロアリール、又は置換若しくは非置換の5～15員のヘテロアリール-C₁～C₆アルキルである、前記実施形態（1）又は（2）に記載の化合物若しくはその塩、又はそれらの溶媒和物。
（4） R⁴が、水素、置換若しくは非置換のC₁～C₆アルキル、置換若しくは非置換のC₂～C₆アルケニル、置換若しくは非置換のC₂～C₆アルキニル、置換若しくは非置換のC₃～C₆シクロアルキル、置換若しくは非置換のC₄～C₆シクロアルケニル、置換若しくは非置換のC₄～C₆シクロアルキニル、置換若しくは非置換の3～6員のヘテロシクロアルキル、置換若しくは非置換のC₇～C₁₁シクロアルキルアルキル、置換若しくは非置換の3～6員のヘテロシクロアルキル-C₁～C₆アルキル、置換若しくは非置換のC₆～C₁₈アリール、置換若しくは非置換のC₇～C₂₀アリールアルキル、置換若しくは非置換の5～15員のヘテロアリール、置換若しくは非置換の5～15員のヘテロアリール-C₁～C₆アルキル、又は置換若しくは非置換のC₁～C₂₀アシルである、前記実施形態（1）～（3）のいずれかに記載の化合物若しくはその塩、又はそれらの溶媒和物。
（5） R¹が、エトキシルであり、
R²が水素であり、
R³が、メチルであり、
R⁴が、メチルである、
前記実施形態（1）～（4）のいずれかに記載の化合物若しくはその塩、又はそれらの溶媒和物。
（6） 前記実施形態（1）～（5）のいずれかに記載の化合物若しくはその塩、又はそれらの溶媒和物の製造方法であって、
式（I）で表される化合物を産生する能力を有するペニシリウム属菌BF-0343株（受託番号NITE P-02833）又はその変異株である微生物を培地中で培養して、式（I）で表される化合物を該培地中に蓄積させる、化合物蓄積工程；
化合物蓄積工程で得られた式（I）で表される化合物を前記微生物の培養物から精製する、化合物精製工程；
を含む、前記方法。
（7） 前記実施形態（1）～（5）のいずれかに記載の式（I）で表される化合物若しくはその塩、又はそれらの溶媒和物を産生する能力を有するペニシリウム属菌BF-0343株（受託番号NITE P-02833）又はその変異株である微生物。
（8） 前記実施形態（1）～（5）のいずれかに記載の化合物若しくはその製薬上許容される塩、又はそれらの製薬上許容される溶媒和物を有効成分として含む、骨代謝阻害剤。
（9） 前記実施形態（1）～（5）のいずれかに記載の化合物若しくはその製薬上許容される塩、又はそれらの製薬上許容される溶媒和物を有効成分として含む医薬。
（10） 骨代謝異常に起因する1種以上の症状、疾患若しくは障害の予防又は治療に使用するための、前記実施形態（9）に記載の医薬。
（11） 骨代謝異常に起因する1種以上の症状、疾患若しくは障害が、進行性骨化性線維異形成症（FOP）又は外傷性異所性骨化である、前記実施形態（10）に記載の医薬。
（12） 前記実施形態（1）～（5）のいずれかに記載の化合物若しくはその製薬上許容される塩、又はそれらの製薬上許容される溶媒和物と、1種以上の製薬上許容される担体とを含む医薬組成物。
（13） 骨代謝異常に起因する1種以上の症状、疾患若しくは障害の予防又は治療に使用するための、前記実施形態（12）に記載の医薬組成物。
（14） 骨代謝異常に起因する1種以上の症状、疾患若しくは障害が、進行性骨化性線維異形成症（FOP）又は外傷性異所性骨化である、前記実施形態（13）に記載の医薬。 That is, the present invention includes the following aspects and embodiments.
(1) Formula (I):

[In the formula,
R ¹ is hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cyclo Alkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy, substituted or unsubstituted heterocyclo alkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted arylalkenyloxy, substituted or unsubstituted heteroaryloxy, substituted or unsubstituted heteroarylalkyloxy, substituted or unsubstituted substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, substituted or unsubstituted acyl, substituted or unsubstituted acyloxy, or substituted or unsubstituted amino;
R ² and R ³ are independently of each other hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted hetero cycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy , substituted or unsubstituted heterocycloalkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted arylalkenyloxy, substituted or unsubstituted heteroaryloxy, substituted or unsubstituted heteroarylalkyloxy, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, substituted or unsubstituted acyl, substituted or unsubstituted acyloxy, or substituted or unsubstituted amino;
^R4 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl , substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted hetero aryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, or substituted or unsubstituted acyl. ]
A compound represented by or a salt thereof, or a solvate thereof.
(2) R ¹ is hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- to 6-membered heterocycloalkoxy, substituted or unsubstituted substituted or unsubstituted C ₆ -C ₁₈ aryloxy, substituted or unsubstituted C ₇ -C ₂₀ arylalkyloxy, substituted or unsubstituted C ₇ -C ₂₀ arylalkenyloxy, substituted or unsubstituted 5- to 15-membered heteroaryl The compound according to embodiment (1) above, which is oxy, substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyloxy, or substituted or unsubstituted C ₁ -C ₂₀ acyloxy, or salts or solvates thereof;
(3) R ² is hydrogen and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted C4- _C6 cycloalkynyl, substituted or unsubstituted _3-6 membered heterocycloalkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3- to 6-membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted _C6 - _C18 aryl, substituted or unsubstituted _C7 - _C20 arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, or substituted or unsubstituted 5-15 membered heteroaryl- _C1 - _C6 alkyl , the compound according to the above embodiment (1) or (2) or a salt thereof, or a solvate thereof.
(4) R ⁴ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3- to 6-membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted _C6 - _C18 aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyl, or substituted or unsubstituted C ₁ - The compound or salt or solvate thereof according to any of the above embodiments (1) to (3), which is C ₂₀ acyl.
(5) ^R1 is ethoxyl;
^R2 is hydrogen,
R ³ is methyl,
R ⁴ is methyl,
The compound according to any one of Embodiments (1) to (4), a salt thereof, or a solvate thereof.
(6) A method for producing a compound or a salt thereof, or a solvate thereof according to any one of Embodiments (1) to (5),
Penicillium strain BF-0343 (accession number NITE P-02833) having the ability to produce the compound represented by formula (I) or a microorganism that is a mutant strain thereof is cultured in a medium, a compound accumulation step, in which the represented compound accumulates in said medium;
A compound purification step of purifying the compound represented by formula (I) obtained in the compound accumulation step from the culture of the microorganism;
The above method, comprising
(7) Penicillium spp. BF-0343 having the ability to produce the compound represented by formula (I) according to any one of the above embodiments (1) to (5), a salt thereof, or a solvate thereof A microorganism that is a strain (accession number NITE P-02833) or a variant thereof.
(8) A bone metabolism inhibitor comprising the compound according to any one of the above embodiments (1) to (5) or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient .
(9) A medicament containing the compound according to any one of the above embodiments (1) to (5), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient.
(10) The medicament according to the above embodiment (9), for use in preventing or treating one or more symptoms, diseases or disorders caused by abnormal bone metabolism.
(11) to the above embodiment (10), wherein the one or more symptoms, diseases or disorders caused by abnormal bone metabolism are fibrodysplasia ossificans progressis (FOP) or traumatic heterotopic ossification; Medication as indicated.
(12) the compound according to any one of the above embodiments (1) to (5) or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more pharmaceutically acceptable and a carrier.
(13) The pharmaceutical composition according to embodiment (12), for use in preventing or treating one or more symptoms, diseases or disorders caused by abnormal bone metabolism.
(14) to the above embodiment (13), wherein the one or more symptoms, diseases or disorders caused by abnormal bone metabolism is fibrodysplasia ossificans progressis (FOP) or traumatic heterotopic ossification; Medication as indicated.

According to one aspect of the present invention, it is possible to provide novel compounds having bone metabolism inhibitory activity, which are highly pharmacologically active and safe.
Problems, configurations, and effects other than those described above will be clarified by the following description of the embodiments.

Preferred embodiments of the present invention are described in detail below.

<1. New compound>
As used herein, "alkyl" means a straight or branched chain saturated aliphatic hydrocarbon group containing the specified number of carbon atoms. For example, "C ₁ -C ₆ alkyl" means a straight or branched chain saturated aliphatic hydrocarbon group containing at least 1 and at most 6 carbon atoms. Suitable alkyls are straight or branched chains such as, but not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl and n-hexyl. Mention may be made of branched C ₁ -C ₆ alkyl.

As used herein, "alkenyl" means a group in which one or more CC single bonds of the above alkyl are replaced with double bonds. Suitable alkenyls include, but are not limited to, vinyl, 1-propenyl, allyl, 1-methylethenyl (isopropenyl), 1-butenyl, 2-butenyl, 3-butenyl, 1-methyl-2-propenyl, 2 -Methyl-2-propenyl, 1-methyl-1-propenyl, 2-methyl-1-propenyl, 1-pentenyl and 1-hexenyl, which may be linear or branched C2 _{- C6} alkenyls. .

As used herein, "alkynyl" means a group in which one or more CC single bonds of the above alkyl are replaced with triple bonds. Suitable alkynyls include, but are not limited to ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-methyl-2-propynyl, 1-pentynyl and 1-hexynyl. and straight or branched chain C ₂ -C ₆ alkynyl.

As used herein, "cycloalkyl" means a cycloaliphatic alkyl containing the specified number of carbon atoms. For example, " _C3 - _C6 cycloalkyl" means a cyclic hydrocarbon group containing at least 3 and at most 6 carbon atoms. Suitable cycloalkyls include, but are not limited to, C ₃ -C ₆ cycloalkyls such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, "cycloalkenyl" means a group in which one or more CC single bonds of the above cycloalkyl are replaced with double bonds. Suitable cycloalkenyls include, but are not limited to, C ₄ -C ₆ cycloalkenyls such as cyclobutenyl, cyclopentenyl and cyclohexenyl.

As used herein, "cycloalkynyl" means a group in which one or more CC single bonds of the above cycloalkyl are replaced with triple bonds. Suitable cycloalkynyls include, but are not limited to, _C4 - _C6 cycloalkynyls such as cyclobutynyl, cyclopentynyl and cyclohexynyl.

As used herein, "heterocycloalkyl" means that one or more carbon atoms of said cycloalkyl, cycloalkenyl or cycloalkynyl are each independently selected from nitrogen (N), sulfur (S) and oxygen (O) means a group substituted with one or more heteroatoms. In this case, substitution by N or S includes substitution of N-oxides or S by oxides or dioxides, respectively. Suitable heterocycloalkyls include, but are not limited to pyrrolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothiopyranyl, piperidinyl, morpholinyl, thiomorpholinyl and piperazinyl. Mention may be made of ~6-membered heterocycloalkyl.

As used herein, "cycloalkylalkyl" means a group in which one hydrogen atom of the above alkyl, alkenyl or alkynyl is replaced with the above cycloalkyl, cycloalkenyl or cycloalkynyl. Suitable cycloalkylalkyls include, but are not limited to, C ₇ -C ₁₁ cycloalkylalkyls such as cyclohexylmethyl and cyclohexenylmethyl.

As used herein, "heterocycloalkylalkyl" means a group in which one hydrogen atom of the above alkyl, alkenyl or alkynyl is substituted with the above heterocycloalkyl. Suitable heterocycloalkylalkyls include, but are not limited to, 3- to 6-membered heterocycloalkyl-C ₁ -C ₆ alkyls.

As used herein, "alkoxy" and "alkoxyl" mean a group in which a hydrogen atom of hydroxyl is substituted with the aforementioned alkyl, alkenyl or alkynyl. Suitable alkoxy and alkoxyl are C ₁ -C ₆ alkoxy or C Mention may be made of ₁ - _C6 alkoxyls.

As used herein, "cycloalkoxy" and "cycloalkoxyl" mean a group in which a hydroxyl hydrogen atom is replaced with the above cycloalkyl, cycloalkenyl or cycloalkynyl. Suitable cycloalkoxy and cycloalkoxyl include, but are not limited to, C ₃ Mention may be made of _-C6 cycloalkoxy or _C3 - _C6 cycloalkoxyl.

As used herein, "heterocycloalkoxy" and "heterocycloalkoxyl" mean a group in which a hydrogen atom of hydroxyl is substituted with the above heterocycloalkyl. Suitable heterocycloalkoxy and heterocycloalkoxyl include, but are not limited to, 3-6 membered heterocycloalkoxy or 3-6 membered heterocycloalkoxyl.

As used herein, "aryl" means an aromatic ring group. Suitable aryls include, but are not limited to, C ₆ -C ₁₈ aryls such as phenyl, biphenyl, terphenyl, naphthyl and anthracenyl.

As used herein, "arylalkyl" means a group in which one hydrogen atom of the above alkyl, alkenyl or alkynyl is substituted with the above aryl. Suitable arylalkyls include, but are not limited to, _C7 - _C20 arylalkyls such as benzyl, 1-phenethyl, 2-phenethyl, biphenylmethyl, terphenylmethyl and styryl.

As used herein, “heteroaryl” means a group in which one or more carbon atoms of said aryl are each independently substituted with one or more heteroatoms selected from N, S and O. In this case, substitution by N or S includes substitution of N-oxides or S by oxides or dioxides, respectively. Suitable heteroaryl include, but are not limited to, furanyl, thienyl (thiophenyl), pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, isothiazolyl, pyridyl, pyridazinyl, pyrazinyl, Mention may be made of 5- to 15-membered heteroaryls such as pyrimidinyl, quinolinyl, isoquinolinyl and indolyl.

As used herein, "heteroarylalkyl" means a group in which one hydrogen atom of the above alkyl, alkenyl or alkynyl is replaced with the above heteroaryl. Suitable heteroarylalkyls include, but are not limited to, 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyls such as pyridylmethyl.

As used herein, "aryloxy" means a group in which a hydroxyl hydrogen atom is substituted with the above aryl. Suitable aryloxy include, but are not limited to, C ₆ -C ₁₈ aryloxy such as phenoxy, biphenyloxy, naphthyloxy and anthryloxy (anthracenyloxy).

As used herein, "arylalkyloxy" means a group in which a hydrogen atom of hydroxyl is substituted with the above arylalkyl. Suitable arylalkyloxy include, but are not limited to, _C7 - _C20 arylalkyloxy such as benzyloxy, 1-phenethyloxy, 2-phenethyloxy and styryloxy.

As used herein, "heteroaryloxy" means a group in which a hydroxyl hydrogen atom is substituted with the above heteroaryl. Suitable heteroaryloxy include, but are not limited to, furanyloxy, thienyloxy (thiophenyloxy), pyrrolyloxy, imidazolyloxy, pyrazolyloxy, triazolyloxy, tetrazolyloxy, thiazolyloxy, oxazolyloxy, iso 5 to 15 of oxazolyloxy, oxadiazolyloxy, thiadiazolyloxy, isothiazolyloxy, pyridyloxy, pyridazinyloxy, pyrazinyloxy, pyrimidinyloxy, quinolinyloxy, isoquinolinyloxy and indolyloxy; membered heteroaryloxy.

As used herein, "heteroarylalkyloxy" means a group in which a hydroxyl hydrogen atom is substituted with the above heteroarylalkyl. Suitable heteroarylalkyloxy include, but are not limited to, 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyloxy.

As used herein, "acyl" means a group in which a monovalent group selected from the groups described above and carbonyl are linked. Suitable acyl are C1 _- C20 acyl including, but not limited to, _C1 - _C5 aliphatic acyl such as formyl, _acetyl and propionyl, and _C7 - _C20 aromatic acyl such as benzoyl. can be mentioned.

Each of the groups described above can be independently unsubstituted or further substituted with one or more monovalent groups described above.

As used herein, "halogen" or "halo" means fluorine (F), chlorine (Cl), bromine (Br) or iodine (I).

で表される化合物若しくはその塩、又はそれらの溶媒和物に関する。 One aspect of the invention is a compound of formula (I):

The compound represented by or a salt thereof, or a solvate thereof.

The present inventors have found that a novel Penicillium bacterium produces in its culture medium a compound having inhibitory activity on BMP signaling. The present inventors discovered a novel compound BF-0343 having bone metabolism inhibitory activity from the culture solution. Compound BF-0343 is a model test system for inhibition of bone morphogenetic protein (BMP) signal transduction involved in bone metabolism. It has a high bone metabolism inhibitory activity that greatly exceeds the compounds described in . The compound represented by Formula (I) of this embodiment includes the natural organic compound BF-0343 and its analogues. Therefore, the compound represented by formula (I) of this embodiment can exhibit high bone metabolism inhibitory activity. In addition, the compound represented by formula (I) of the present invention has a different structure from the compounds of the prior art exemplified above. There is a possibility that a therapeutic effect can be expressed.

In formula (I), R ¹ is hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkyl alkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy, substituted or unsubstituted heterocycloalkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted arylalkenyloxy, substituted or unsubstituted heteroaryloxy, substituted or unsubstituted heteroaryl alkyloxy, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, substituted or unsubstituted acyl, substituted or unsubstituted acyloxy, or substituted or unsubstituted amino. R ¹ is hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5-15 membered heteroaryl-C _{1 -C6} alkyl, substituted or unsubstituted C1 _{- C6} alkoxy, substituted or unsubstituted C3 _{- C6} cycloalkoxy, substituted or unsubstituted 3-6 membered heterocycloalkoxy, substituted or unsubstituted C ₆ - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, _substituted or unsubstituted C7- _C20 arylalkenyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyloxy, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, substituted or unsubstituted C ₃ -C ₆ cycloalkoxycarbonyl, substituted or unsubstituted preferably substituted C ₁ -C ₂₀ acyl, substituted or unsubstituted C ₁ -C ₂₀ acyloxy, or substituted or unsubstituted amino; hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- to 6-membered heterocycloalkoxy, substituted or unsubstituted C ₆ -C ₁₈ aryloxy, substituted or unsubstituted C ₇ -C ₂₀ arylalkyloxy , substituted or unsubstituted C ₇ -C ₂₀ arylalkenyloxy, substituted or unsubstituted 5- to 15-membered heteroaryloxy, substituted or unsubstituted 5- to 15-membered heteroaryl- more preferably C ₁ -C ₆ alkyloxy, or substituted or unsubstituted C ₁ -C ₂₀ acyloxy; substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cyclo alkoxy, substituted or unsubstituted 3- to 6-membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted _C7- more preferably C ₂₀ arylalkenyloxy, substituted or unsubstituted 5- to 15-membered heteroaryloxy, or substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyloxy; substituted or Unsubstituted C ₁ -C ₆ alkoxy is even more preferred; methoxyl, ethoxyl, propoxyl, butoxyl, pentoxyl or hexoxyl are particularly preferred; ethoxyl is especially preferred. When R ¹ is the group exemplified above, the compound represented by formula (I) of this embodiment can exhibit low cytotoxicity and high bone metabolism inhibitory activity.

In formula (I), R ² and R ³ are independently of each other hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl , substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy, substituted or unsubstituted heterocycloalkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted arylalkenyloxy, substituted or unsubstituted heteroaryloxy , substituted or unsubstituted heteroarylalkyloxy, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, substituted or unsubstituted acyl, substituted or unsubstituted acyloxy, or substituted or unsubstituted amino be. R ² and R ³ are independently of each other hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C _{2 -C6} alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted C3 _{- C6} cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _{C4 -C6} cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl- _{C1 -C6} alkyl, substituted or unsubstituted _C6 - _C18 aryl, substituted or unsubstituted _C7 - _C20 arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5- 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3-6 membered heterocyclo alkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted C7 _{- C20} arylalkyloxy, substituted or unsubstituted C7- _C20 arylalkenyloxy, substituted or unsubstituted _5- 15-membered heteroaryloxy, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyloxy, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, substituted or unsubstituted C ₃ -C ₆ is preferably cycloalkoxycarbonyl, substituted or unsubstituted C ₁ -C ₂₀ acyl, substituted or unsubstituted C ₁ -C ₂₀ acyloxy, or substituted or unsubstituted amino; R ² is hydrogen; and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C4 _{- C6} cycloalkenyl, substituted or unsubstituted C4 _{- C6} cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C ₇ -C ₁₁ Cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl , substituted or unsubstituted 5- to 15-membered heteroaryl, or substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl; R ² is hydrogen, and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₇ -C ₁₁ Cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, or substituted or unsubstituted 5-15 membered hetero more preferably aryl-C ₁ -C ₆ alkyl; even more preferably R ² is hydrogen and ^{R 3} is substituted or unsubstituted C ₁ -C ₆ alkyl; , hydrogen and R ³ is methyl, ethyl, propyl, butyl, pentyl or hexyl; particularly preferably R ² is hydrogen and R ³ is methyl preferable. When R ² and R ³ are the groups exemplified above, the compound represented by formula (I) of this embodiment can exhibit low cytotoxicity and high bone metabolism inhibitory activity.

In formula (I), ^R4 is hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, or substituted or unsubstituted acyl. R ⁴ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ - C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C ₇ -C ₁₁ cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ - C ₂₀ arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl , substituted or unsubstituted C ₃ -C ₆ cycloalkoxycarbonyl, or substituted or unsubstituted C ₁ -C ₂₀ acyl; hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted substituted or unsubstituted _C4 - _C6 cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, _substituted or unsubstituted 3-6 membered heterocyclo Alkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted more preferably substituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, or substituted or unsubstituted C ₁ -C ₂₀ acyl; substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3- to 6-membered heterocycloalkyl- _C1 - _C6 more preferably alkyl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, or substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl; substituted or unsubstituted C ₁ - _C6 alkyl is even more preferred; methyl, ethyl, propyl, butyl, pentyl or hexyl is particularly preferred; methyl is especially preferred. When R ⁴ is the group exemplified above, the compound represented by formula (I) of this embodiment can exhibit low cytotoxicity and high bone metabolism inhibitory activity.

In formula (I), when said groups are substituted, said substituents are each independently halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloalkenyl, substituted or unsubstituted cycloalkynyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted cycloalkylalkyl, substituted or unsubstituted heterocycloalkylalkyl, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroarylalkyl, substituted or unsubstituted substituted or unsubstituted alkoxy, substituted or unsubstituted cycloalkoxy, substituted or unsubstituted heterocycloalkoxy, substituted or unsubstituted aryloxy, substituted or unsubstituted arylalkyloxy, substituted or unsubstituted heteroaryloxy, substituted or unsubstituted substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted cycloalkoxycarbonyl, substituted or unsubstituted acyl, substituted or unsubstituted acyloxy, substituted or unsubstituted amino, and oxo (C= O) is preferably at least one monovalent or divalent group selected from the group consisting of halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C _{1 -C6} alkyl, substituted or unsubstituted _C2 -C6 alkenyl, substituted or unsubstituted C2 _{- C6} alkynyl, substituted or unsubstituted _{C3 -C6 cycloalkyl} , substituted or unsubstituted _{C4- C6} cycloalkenyl, substituted or unsubstituted C4 _{- C6 cycloalkynyl} , substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5-15 heteroaryl, substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- to 6-membered heterocycloalkoxy, substituted or unsubstituted C ₆ -C ₁₈ aryloxy , substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted 5-15 membered heteroaryl- _C1 - _C6 alkyloxy, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, substituted or unsubstituted C ₃ -C ₆ cycloalkoxycarbonyl, substituted or unsubstituted C ₁ -C ₂₀ acyl, substituted or unsubstituted C ₁ -C ₂₀ acyloxy , substituted or unsubstituted amino, and oxo (C═O), more preferably at least one monovalent or divalent group selected from the group consisting of hydroxyl, substituted or unsubstituted C ₁ to _C6 alkoxy, substituted or unsubstituted C3 _{- C6} cycloalkoxy, substituted or unsubstituted 3-6 membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _{C7 -C20} arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted 5-15 membered heteroaryl- _C1 - _C6 alkyloxy, and substituted or unsubstituted _C1 More preferably, it is at least one monovalent group selected from the group consisting of _-C20 acyloxy, particularly preferably hydroxyl. When said monovalent group is substituted, said substituent is preferably further selected from said monovalent group or divalent group, and further selected from said unsubstituted monovalent group or divalent group. is more preferred.

Compounds represented by formula (I) can include compounds defined by any combination of R ¹ , R ² , R ³ and R ⁴ exemplified above.

Preferably, the compound of formula (I) is
R ¹ is hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ -C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5-15 membered heteroaryl-C _{1 -C6} alkyl, substituted or unsubstituted C1 _{- C6} alkoxy, substituted or unsubstituted C3 _{- C6} cycloalkoxy, substituted or unsubstituted 3-6 membered heterocycloalkoxy, substituted or unsubstituted C ₆ - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, _substituted or unsubstituted C7- _C20 arylalkenyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyloxy, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, substituted or unsubstituted C ₃ -C ₆ cycloalkoxycarbonyl, substituted or unsubstituted substituted C ₁ -C ₂₀ acyl, substituted or unsubstituted C ₁ -C ₂₀ acyloxy, or substituted or unsubstituted amino;
R ² and R ³ are independently of each other hydrogen, halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C _{2 -C6} alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted C3 _{- C6} cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _{C4 -C6} cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl- _{C1 -C6} alkyl, substituted or unsubstituted _C6 - _C18 aryl, substituted or unsubstituted _C7 - _C20 arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5- 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3-6 membered heterocyclo alkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted C7 _{- C20} arylalkyloxy, substituted or unsubstituted C7- _C20 arylalkenyloxy, substituted or unsubstituted _5- 15-membered heteroaryloxy, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyloxy, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, substituted or unsubstituted C ₃ -C ₆ cycloalkoxycarbonyl, substituted or unsubstituted C ₁ -C ₂₀ acyl, substituted or unsubstituted C ₁ -C ₂₀ acyloxy, or substituted or unsubstituted amino;
R ⁴ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ - C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C ₇ -C ₁₁ cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ - C ₂₀ arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl , substituted or unsubstituted _C3 - _C6 cycloalkoxycarbonyl, or substituted or unsubstituted C1 _{- C20} acyl;
When said groups are substituted, said substituents are each independently halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _C4 - _C6 cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, substituted or unsubstituted _3-6 membered hetero Cycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- 6-membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, _{substituted or unsubstituted C 3 -C 6 cycloalkoxycarbonyl} , substituted or unsubstituted at least one monovalent or divalent group selected from the group consisting of C1- _C20 acyl, substituted or unsubstituted _C1 - _C20 acyloxy, substituted or unsubstituted amino _, and oxo (C=O); is the base. When said monovalent group is substituted, said substituent is preferably further selected from said monovalent group or divalent group, and further selected from said unsubstituted monovalent group or divalent group. is more preferred.

More preferably, the compound of formula (I) is
R ¹ is hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3-6 membered heterocycloalkoxy, substituted or unsubstituted C ₆ - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, _substituted or unsubstituted C7- _C20 arylalkenyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyloxy, or substituted or unsubstituted C ₁ -C ₂₀ acyloxy;
R ² is hydrogen and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted C4 _{- C6} cycloalkenyl, substituted or unsubstituted C4- _C6 cycloalkynyl, substituted or unsubstituted _3-6 membered heterocyclo Alkyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3- to 6-membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted _C6 - _C18 aryl, substituted or unsubstituted _C7 - _C20 arylalkyl, substituted or unsubstituted 5-15 membered heteroaryl, or substituted or unsubstituted 5-15 membered heteroaryl- _C1 - _C6 alkyl;
R ⁴ is hydrogen, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₃ - C ₆ cycloalkyl, substituted or unsubstituted C ₄ -C ₆ cycloalkenyl, substituted or unsubstituted C ₄ -C ₆ cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C ₇ -C ₁₁ cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ - C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, or substituted or unsubstituted C ₁ -C ₂₀ acyl is;
When said groups are substituted, said substituents are each independently halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _C4 - _C6 cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, substituted or unsubstituted _3-6 membered hetero Cycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- 6-membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, _{substituted or unsubstituted C 3 -C 6 cycloalkoxycarbonyl} , substituted or unsubstituted at least one monovalent or divalent group selected from the group consisting of C1- _C20 acyl, substituted or unsubstituted _C1 - _C20 acyloxy, substituted or unsubstituted amino _, and oxo (C=O); is the base. When said monovalent group is substituted, said substituent is preferably further selected from said monovalent group or divalent group, and further selected from said unsubstituted monovalent group or divalent group. is more preferred.

More preferably, the compound of formula (I) is
R ¹ is substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- to 6-membered heterocycloalkoxy, substituted or unsubstituted C ₆ - C ₁₈ aryloxy, substituted or unsubstituted C ₇ -C ₂₀ arylalkyloxy, substituted or unsubstituted C ₇ -C ₂₀ arylalkenyloxy, substituted or unsubstituted 5- to 15-membered heteroaryloxy, or substituted or unsubstituted 5-15 membered heteroaryl-C ₁ -C ₆ alkyloxy;
R ² is hydrogen and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted _C7 - _C11 cycloalkylalkyl, substituted or unsubstituted 3- to 6-membered heterocycloalkyl- _C1 - _C6 alkyl, substituted or unsubstituted _C7 - _C20 arylalkyl, or substituted or is unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl;
R ⁴ is substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted C ₂ -C ₆ alkenyl, substituted or unsubstituted C ₂ -C ₆ alkynyl, substituted or unsubstituted C ₇ -C ₁₁ Cycloalkylalkyl, substituted or unsubstituted 3-6 membered heterocycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, or substituted or unsubstituted 5-15 membered hetero aryl-C ₁ -C ₆ alkyl;
When said groups are substituted, said substituents are each independently halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _C4 - _C6 cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, substituted or unsubstituted _3-6 membered hetero Cycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- 6-membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, _{substituted or unsubstituted C 3 -C 6 cycloalkoxycarbonyl} , substituted or unsubstituted at least one monovalent or divalent group selected from the group consisting of C1- _C20 acyl, substituted or unsubstituted _C1 - _C20 acyloxy, substituted or unsubstituted amino _, and oxo (C=O); is the base. When said monovalent group is substituted, said substituent is preferably further selected from said monovalent group or divalent group, and further selected from said unsubstituted monovalent group or divalent group. is more preferred.

Even more preferably, the compound of formula (I) is
R ¹ is substituted or unsubstituted C ₁ -C ₆ alkoxy;
R ² is hydrogen and R ³ is substituted or unsubstituted C ₁ -C ₆ alkyl;
R ⁴ is substituted or unsubstituted C ₁ -C ₆ alkyl;
When said groups are substituted, said substituents are each independently halogen (fluorine, chlorine, bromine or iodine), cyano, nitro, hydroxyl, substituted or unsubstituted C ₁ -C ₆ alkyl, substituted or unsubstituted _C2 - _C6 alkenyl, substituted or unsubstituted _C2 - _C6 alkynyl, substituted or unsubstituted _C3 - _C6 cycloalkyl, substituted or unsubstituted _C4 - _C6 cycloalkenyl, substituted or unsubstituted _C4 - _C6 cycloalkynyl, substituted or unsubstituted 3-6 membered heterocycloalkyl, substituted or unsubstituted C7- _C11 cycloalkylalkyl, substituted or unsubstituted _3-6 membered hetero Cycloalkyl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₆ -C ₁₈ aryl, substituted or unsubstituted C ₇ -C ₂₀ arylalkyl, substituted or unsubstituted 5- to 15-membered heteroaryl, substituted or unsubstituted unsubstituted 5- to 15-membered heteroaryl-C ₁ -C ₆ alkyl, substituted or unsubstituted C ₁ -C ₆ alkoxy, substituted or unsubstituted C ₃ -C ₆ cycloalkoxy, substituted or unsubstituted 3- 6-membered heterocycloalkoxy, substituted or unsubstituted _C6 - _C18 aryloxy, substituted or unsubstituted _C7 - _C20 arylalkyloxy, substituted or unsubstituted 5-15 membered heteroaryloxy, substituted or unsubstituted substituted or unsubstituted C ₁ -C ₆ alkoxycarbonyl, _{substituted or unsubstituted C 3 -C 6 cycloalkoxycarbonyl} , substituted or unsubstituted at least one monovalent or divalent group selected from the group consisting of C1- _C20 acyl, substituted or unsubstituted _C1 - _C20 acyloxy, substituted or unsubstituted amino _, and oxo (C=O); is the base. When said monovalent group is substituted, said substituent is preferably further selected from said monovalent group or divalent group, and further selected from said unsubstituted monovalent group or divalent group. is more preferred.

Particularly preferably, the compound of formula (I) is
R ¹ is methoxyl, ethoxyl, propoxyl, butoxyl, pentoxyl or hexoxyl;
^R2 is hydrogen and ^R3 is methyl, ethyl, propyl, butyl, pentyl or hexyl;
^R4 is methyl, ethyl, propyl, butyl, pentyl or hexyl.

Particularly preferably, the compound of formula (I) is
R ¹ is ethoxyl;
R ² is hydrogen,
R ³ is methyl;
^R4 is methyl.

Especially particularly preferred compounds of formula (I) are:
5-(1-ethoxypropan-2-yl)-5-hydroxy-4-methoxyfuran-2(5H)-one (BF-0343);
is. When the compound represented by formula (I) of this embodiment is the compound described above, the compound can exhibit high bone metabolism inhibitory activity.

The compound represented by Formula (I) in one embodiment of the present invention includes not only the compound itself but also salts thereof. Salts of the compound represented by Formula (I) of one embodiment of the present invention are not limited, but examples include sodium ion, potassium ion, calcium ion, magnesium ion, or substituted or unsubstituted ammonium ion. or inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, carbonic acid or phosphoric acid, or formic acid, acetic acid, maleic acid, fumaric acid, benzoic acid, ascorbic acid, succinic acid, bis methylenesalicylic acid, methanesulfonic acid, ethanedisulfonic acid, propionic acid, tartaric acid, salicylic acid, citric acid, gluconic acid, aspartic acid, stearic acid, palmitic acid, itaconic acid, glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid Salts with organic acid anions such as , cyclohexylsulfamic acid, methanesulphonic acid, ethanesulphonic acid, isethionic acid, p-toluenesulphonic acid or naphthalenesulphonic acid are preferred. Even when the compound represented by formula (I) is in the above salt form, it can exhibit high bone metabolism inhibitory activity.

The compound represented by formula (I) in one embodiment of the present invention includes not only the compound itself but also solvates of the compound or a salt thereof. Solvents capable of forming solvates with the compounds or salts thereof include, but are not limited to, water or lower alcohols such as methanol, ethanol or 2-propanol (isopropyl alcohol). -6 carbon atoms), higher alcohols (e.g. alcohols with 7 or more carbon atoms such as 1-heptanol or 1-octanol), dimethyl sulfoxide (DMSO), acetic acid, ethanolamine or ethyl acetate Organic solvents such as are preferred. Even when the compound represented by formula (I) or a salt thereof is in the form of a solvate with the solvent, it can exhibit high bone metabolism inhibitory activity.

The compound represented by formula (I) in one embodiment of the present invention includes not only the compound itself but also protected forms thereof. As used herein, "protected form" means a form in which a protecting group is introduced to one or more functional groups (eg, amino group, hydroxyl group or carboxylic acid group). In this specification, protected forms of the compounds represented by the above formulas are sometimes referred to as protected derivatives of the compounds represented by the above formulas. Also, as used herein, a "protecting group" is a group introduced into a specific functional group in order to prevent the progress of an undesired reaction, and is quantitatively removed under specific reaction conditions and It means a group which is substantially stable, ie, reaction inert, under reaction conditions other than . The protective group capable of forming a protected form of the compound is not limited, but for example, in the case of an amino group protective group, t-butoxycarbonyl (Boc), 2-bromobenzyloxycarbonyl (BrZ), or When 9-fluorenylmethoxycarbonyl (Fmoc) is a hydroxyl-protecting group, silyl (e.g., t-butyldimethylsilyl (TBS), triisopropylsilyl (TIPS) or tert-butyldiphenylsilyl (TBDPS)), or when alkoxy (e.g. methoxymethoxy (MOM) or methoxy (Me)) is a protecting group for a carboxylic acid group, an alkyl ester (e.g. methyl, ethyl or isopropyl ester), an arylalkyl ester (e.g. benzyl ester), or an amide (eg amides with oxazolidinones) are respectively preferred. Protection and deprotection with the protecting group can be appropriately carried out by those skilled in the art based on known reaction conditions. Even when the compound represented by formula (I) of one embodiment of the present invention is in a form protected by the protective group, it may exhibit high bone metabolism inhibitory activity.

When a compound represented by Formula (I) in one aspect of the present invention has one or more tautomers, said compound also includes the individual tautomeric forms of said compound.

In addition, when the compound represented by formula (I) of one embodiment of the present invention has one or more stereocenters (chiral centers), the compound may be the individual enantiomers and diastereomers of the compound, and Mixtures thereof such as racemates are also included.

By having the above characteristics, the compound represented by formula (I) of one embodiment of the present invention can exhibit high bone metabolism inhibitory activity.

<2. Method for producing novel compounds by culture means>
The present inventors have found that the novel Penicillium bacterium produces in its culture medium the compound BF-0343, which is included in the compound represented by formula (I) of one embodiment of the present invention. Therefore, another aspect of the invention relates to a method of producing a compound of formula (I) according to one aspect of the invention using the novel Penicillium spp.

In this aspect, the method of the invention includes a compound accumulation step and a compound purification step. Each step will be described in detail below.

[2-1. Compound accumulation step]
In this embodiment, the method of the present invention is a microorganism that is a Penicillium spp. in a medium to accumulate the compound represented by formula (I) in the medium.

Penicillium strain BF-0343 is a novel filamentous fungus belonging to the genus Penicillium isolated from soil in Shizuoka Prefecture, Japan. This strain was registered as BF-0343 on November 28, 2018 at the Patent Microorganism Depositary Center, National Institute of Technology and Evaluation (2-5-8-122 Kazusa Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan). room) (accession number NITE P-02833).

In this step, the microorganism used for culturing is preferably Penicillium spp. More preferably, it is a strain. In the present invention, a mutant strain of the genus Penicillium strain BF-0343 means a natural mutant strain or an artificial mutant strain of the genus Penicillium strain BF-0343. The artificial mutant strain of Penicillium spp. BF-0343 strain can be obtained by any artificial mutant production means commonly used in the art. Not only the Penicillium spp. BF-0343 strain itself, but also a mutant strain of the Penicillium spp. A compound of formula (I) can accumulate in the medium. Therefore, by using the microorganism, a large amount of the compound represented by formula (I) can be accumulated in the medium.

The medium used for culturing the microorganism can be appropriately selected based on the properties of the microorganism. The medium usually contains one or more carbon sources and one or more nitrogen sources, and optionally one or more inorganic salts and one or more vitamins. Carbon sources include sugars such as glucose, fructose, maltose, lactose, galactose, dextrin and starch, and vegetable oils such as soybean oil. Nitrogen sources can include polypeptones, yeast extracts, malt extracts, meat extracts, soy flour, cottonseed flour, corn-steve liquor, casein, amino acids, urea, ammonium salts and nitrates. Inorganic salts include cations such as sodium ion, potassium ion, calcium ion, magnesium ion, iron ion, manganese ion, copper ion, cobalt ion or zinc ion, and inorganic salts such as hydrochloric acid, sulfuric acid, nitric acid, carbonic acid or phosphoric acid. Salts with acid anions may be mentioned. The medium used in this step is, for example, potato dextrose agar (PDA) medium (3.9% potato dextrose agar (DIFCO), adjusted to pH 6.0) or potato dextrose broth (PDB) medium (2.4% Potato dextrose broth (DIFCO), 0.1% agar (SSK sales), adjusted to pH 6.0) is preferred. The compound represented by formula (I) is produced by culturing a microorganism that is a Penicillium strain BF-0343 or a mutant strain thereof that has the ability to produce the compound represented by formula (I) in the medium. It can accumulate in the medium.

Microorganism culture may be either solid culture or liquid culture. When culturing the microorganisms on a large scale, liquid culture is preferred. In this case, it is preferable to aerate the medium by shaking the culture vessel or agitating the medium with a propeller or the like, or blowing air with a pump or the like. The air introduced into the medium is preferably sterilized using a sterilizing means such as a sterilizing filter. By culturing the microorganism under the above conditions, the compound represented by formula (I) can be efficiently accumulated in the medium.

When culturing the microorganisms on a large scale, the microorganisms are cultured in advance in a small amount of medium (hereinafter also referred to as "seed culture"), and then the culture obtained by the seed culture is planted in a large volume of medium. It is preferable to culture the bacteria (hereinafter also referred to as “production culture”). In this case, the components of the medium used for seed culture and production culture may be the same or different. By carrying out this step through multiple stages of culture including seed culture and production culture, delay in growth of microorganisms can be substantially suppressed.

In this step, the conditions for culturing the microorganism can be appropriately set based on the properties of the microorganism. The culture temperature is usually in the range of 25-27°C, typically around 25°C. The pH of the medium is usually pH 3-9, typically around pH 5.6. The culture period is usually 3 to 12 days, typically 5 to 10 days, in total for seed culture and production culture when a liquid medium is cultured with shaking. By culturing the microorganism under the above conditions, the compound represented by formula (I) can be efficiently accumulated in the medium.

[2-2. Compound accumulation step]
In this aspect, the method of the present invention includes a compound purification step of purifying the compound represented by formula (I) obtained in the compound accumulation step from the culture of the microorganism.

In this step, as means for purifying the compound represented by formula (I) from the culture of microorganisms, a separation method for organic compounds commonly used in the art can be used. Examples of means for purifying the compound represented by formula (I) from a microbial culture include extraction, filtration, centrifugation, adsorption, recrystallization, distillation, and various types of chromatography. Preferably, this step includes a step of separating the cells from the microbial culture obtained in the compound accumulation step by filtration or centrifugation, a step of extracting the separated cells with an organic solvent such as ethanol or ethyl acetate, and a step of further separating the bacterial cell extract extracted with an organic solvent by means such as solvent extraction or preparative chromatography to obtain the compound represented by formula (I). Various types of chromatography such as adsorption, normal phase or reverse phase partition, or gel filtration can be applied as the preparative chromatography. In the final step, the fractions obtained by preparative chromatography may be further purified by means such as recrystallization or distillation. Each of the above steps may be repeated multiple times under the same or different conditions, if desired. By using said means, compounds of formula (I) can be purified and isolated from cultures of microorganisms.

In the method for producing the compound of the present invention by the culture method of this aspect, the compound represented by formula (I) preferably has the groups R ¹ , R ² , R ³ and R ⁴ exemplified above. In this case, the compound represented by formula (I) includes compound BF-0343 produced by Penicillium strain BF-0343. By using the method of this embodiment, the compound represented by formula (I) corresponding to compound BF-0343 can be efficiently produced.

By the method for producing the compound of the present invention by the culturing method of this aspect having the above characteristics, the compound represented by the formula (I) of one aspect of the present invention, which can be an active ingredient of a medicine, can be produced with high purity and at low cost. Can be provided in large quantities.

Another aspect of the present invention is Penicillium strain BF-0343 (accession number NITE P-02833) having the ability to produce a compound represented by formula (I) of one aspect of the present invention, or a mutant strain thereof. Concerning certain microorganisms. In this embodiment, the compound represented by formula (I) preferably has the groups R ¹ , R ² , R ³ and R ⁴ exemplified above. By using the microorganism of this embodiment, the compound represented by formula (I) corresponding to compound BF-0343 can be produced efficiently.

<3. Pharmaceutical use>
The compound represented by formula (I) of one embodiment of the present invention has high BMP signaling inhibitory activity, but does not substantially show cytotoxicity in the concentration range where BMP signaling inhibitory activity is exhibited. Therefore, one aspect of the present invention comprises a compound represented by formula (I) of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient It relates to BMP signaling inhibitors. In addition, when the compound represented by formula (I) of one embodiment of the present invention is administered to a subject, the subject expresses bone metabolism inhibitory activity through high BMP signaling inhibitory activity and is caused by abnormal bone metabolism. can prevent or treat a particular symptom, disease or disorder of Therefore, another aspect of the present invention is a compound represented by formula (I) of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient. It relates to a bone metabolism inhibitor, medicament or pharmaceutical composition comprising as

The BMP signal transduction inhibitory activity of the compound represented by formula (I) of one embodiment of the present invention is not limited, but is, for example, C2C12 cells, a myoblast cell line established from the muscle regeneration part of growing mice. as a model experimental system (T. Katagiri et al., J. Cell Biol., 1994, vol. 127, p. 1755-1766). When a specific compound is added to the BMP-induced differentiation system of C2C12 cells into osteoblasts, differentiation into osteoblasts, that is, bone metabolism is inhibited, and the expression level of alkaline phosphatase is reduced. Therefore, using alkaline phosphatase activity as an index, BMP signal transduction inhibitory activity against the differentiation induction system from C2C12 cells to osteoblasts, that is, bone metabolism inhibitory activity can be evaluated (T. Fukuda et al., J. Biol. Chem. ., 2009, Vol. 284, p. 7149-7156). In this case, the BMP signaling inhibitory activity of the compound represented by formula (I) of one embodiment of the present invention can be determined, for example, by the following procedure. Mouse muscle-derived C2C12 cells stably transfected with a gene in which the 206th arginine of ALK2 is mutated to histidine (hereinafter also referred to as "C2C12 (R206H) cells") were suspended in a predetermined medium to form a cell suspension. to prepare. A predetermined amount of this cell suspension is seeded into each well of a 96-well microplate. After cell seeding, the microplate is transferred to a carbon dioxide incubator and the cells are cultured for a certain period of time, after which the medium in each well is replaced with a medium containing a predetermined amount of recombinant human BMP-4. A given final concentration of test compound is added to a given well. The cells are then cultured for a period of time. After the culture is completed, the culture supernatant is removed and the cells are washed. Fix the cells using a fixative solution. After washing the cells, 4-nitrophenyl phosphate (4-NPP) solution is dispensed into each well. After performing the enzymatic reaction under predetermined conditions, the absorbance of the solution in each well (for example, absorbance at a wavelength of 405 nm) is measured using a microplate reader. The percent inhibition of BMP signaling for each test compound can be calculated based on the absorbance values obtained. The BMP signaling inhibitory activity of the compound represented by formula (I) of one embodiment of the present invention is based on the inhibition rate of the BMP signaling inhibitory activity determined by the above procedure, and the compound of the present invention that inhibits BMP signaling by 50%. It is preferably expressed as the concentration (ALP-IC ₅₀ ) of the compound represented by Formula (I) in one embodiment. The compound represented by formula (I) of one aspect of the present invention has an ALP-IC ₅₀ value of BMP signaling inhibitory activity determined by the above procedure of usually 50 μM or less, typically 20 μM or less. . When having an ALP-IC ₅₀ value within the above range, the compound represented by formula (I) of one embodiment of the present invention can exhibit high BMP signaling inhibitory activity. In this case, the compound represented by formula (I) of one embodiment of the present invention can exhibit bone metabolism inhibitory activity through high BMP signal transduction inhibitory activity.

The bone metabolism inhibitory activity of the compound represented by formula (I) of one aspect of the present invention can be determined based on the BMP signaling inhibitory activity described above. Alternatively, the bone metabolism inhibitory activity of the compound represented by formula (I) of one embodiment of the present invention is, but not limited to, Western blotting using an antibody against ALK2 or SMAD4 that transmits an ossification signal. or immunostaining, measurement of the amount of osteocalcin released in the culture supernatant, or measurement of responsiveness to parathyroid hormone (T. Katagiri et al., J. Cell Biol ., 1994, Vol. 127, p. 1755-1766).

The safety of the compound represented by formula (I) of one embodiment of the present invention is evaluated by, but not limited to, an MTT evaluation method using thiazolyl blue tetrazolyl bromide (MTT) (Mosmann et al., J Immunol Methods, 1983 2005, vol. 65, p. 55-63), by evaluating the cytotoxicity of the compound. The cytotoxicity of the compound represented by formula (I) of one aspect of the present invention is based on the cell viability determined by the above procedure, and the formula (I) of one aspect of the present invention inhibits cell survival by 50%. It is preferably expressed as the concentration of the compound represented by (MTT-IC ₅₀ ). The compound represented by formula (I) of one aspect of the present invention is used in a concentration range in which the cell viability determined by the above procedure substantially expresses BMP signaling inhibitory activity and/or bone metabolism inhibitory activity, Usually 50% or more, typically 60%. Alternatively, the compound represented by formula (I) of one aspect of the present invention has a cytotoxic MTT-IC ₅₀ value determined by the above procedure of usually 10 μM or more, typically 20 μM or more, particularly 30 μM or more. μM or higher. When the cell viability and/or MTT-IC ₅₀ value are within the above range, the compound represented by formula (I) of one embodiment of the present invention substantially exhibits BMP signaling inhibitory activity and/or bone metabolism inhibitory activity. It can be safely used without substantially exhibiting cytotoxicity within the concentration range in which it is practically expressed.

When the compound represented by formula (I) of one embodiment of the present invention is applied to medical use, the compound represented by formula (I) is not only the compound itself, but also a pharmaceutically acceptable salt of the compound. , and pharmaceutically acceptable solvates thereof. Pharmaceutically acceptable salts and pharmaceutically acceptable solvates thereof of the compound represented by formula (I) of one embodiment of the present invention are not limited, but are exemplified above. Salts or solvates are preferred. When the compound represented by formula (I) is in the form of a pharmaceutically acceptable salt or pharmaceutically acceptable solvate, the bone metabolism inhibitory activity is not substantially reduced, and/or The compounds can be applied for desired pharmaceutical uses without substantially increasing toxicity.

When the compound represented by formula (I) of one embodiment of the present invention is applied to pharmaceutical use, the compound represented by formula (I) includes not only the compound itself but also prodrug forms of the compound. . As used herein, "prodrug" means a compound that is converted in vivo to the parent drug. Prodrug forms of the compounds include, but are not limited to, esters of the hydroxyl group with any carboxylic acid, and amides of the hydroxyl group with any amine, if a hydroxyl group is present. For example, when an amino group is present, amides of the amino group and any carboxylic acid can be mentioned. When the compound represented by formula (I) of one aspect of the present invention is in the prodrug form, the bone metabolism inhibitory activity of the parent drug represented by formula (I) is substantially reduced. and/or without substantially increasing cytotoxicity, pharmacokinetics upon administration of the prodrug form to a subject can be improved.

When the compound represented by formula (I) of one embodiment of the present invention is applied for pharmaceutical use, the compound may be used alone or in combination with one or more pharmaceutically acceptable ingredients. good too. The medicament of this aspect can be formulated into various dosage forms commonly used in the art, depending on the desired method of administration. Therefore, the medicament of this aspect also includes a compound represented by formula (I) of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more and a pharmaceutically acceptable carrier. In addition to the above ingredients, the pharmaceutical composition of this embodiment contains one or more pharmaceutically acceptable vehicles (e.g., solvents such as sterile water or solutions such as physiological saline), excipients, binders, Vehicles, solubilizers, preservatives, stabilizers, swelling agents, lubricants, surfactants, emulsifiers, oily liquids (e.g. vegetable oils), suspending agents, buffers, soothing agents, antioxidants, sweeteners and Flavoring agents and the like may also be included.

A pharmaceutical dosage form containing the compound represented by formula (I) of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient is not particularly limited. , it may be a formulation for use in parenteral administration, or it may be a formulation for use in oral administration. In addition, the dosage form of the medicament of this aspect may be a unit dosage form formulation or a multiple dosage form formulation. Formulations for parenteral administration include injections such as sterile solutions or suspensions in water or other pharmaceutically acceptable vehicles. Ingredients that can be mixed with injections include, but are not limited to, physiological saline, glucose, or other adjuvants (such as D-sorbitol, D-mannitol, D-mannose, or sodium chloride). Vehicles such as isotonic solutions containing, solubilizing agents such as alcohols (e.g. ethanol or benzyl alcohol), polyalcohols (e.g. propylene glycol or polyethylene glycol) or esters (e.g. benzyl benzoate), polysorbate 80 (trademark) or poly Non-ionic surfactants such as oxyethylene hydrogenated castor oil, oily liquids such as sesame oil or soybean oil, buffers such as phosphate buffers or sodium acetate buffers, benzalkonium chloride or procaine hydrochloride. soothing agents, stabilizers such as human serum albumin or polyethylene glycol, preservatives, and antioxidants. The prepared injection is usually filled into a suitable vial (for example, an ampoule) and stored under an appropriate environment until use.

Formulations for oral administration include, for example, tablets, pills, powders, capsules, microcapsules, elixirs, liquids, syrups, slurries and suspensions. Tablets may optionally be formulated as sugar-coated or dissolution-coated sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets or film-coated tablets, or as double- or multi-layered tablet dosage forms. may be formulated.

Ingredients that can be mixed with tablets or capsules include, but are not limited to, water, ethanol, propanol, simple syrup, glucose solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, binders such as gelatin, corn starch, gum tragacanth or gum arabic; excipients such as crystalline cellulose, lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin or silicic acid; dry starch, sodium alginate. , agar powder, laminaran powder, sodium hydrogen carbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, monoglyceride stearate, starch or lactose; absorption enhancers such as quaternary ammonium salts or sodium lauryl sulfate; humectants such as glycerin or starch; adsorbents such as starch, lactose, kaolin, bentonite or colloidal silicic acid; purified talc; Lubricants such as stearates (e.g. magnesium stearate), powdered boric acid or polyethylene glycol; sweetening agents such as sucrose, lactose or saccharin; and flavoring agents such as peppermint, redwood oil or cherry. can be done. When the formulation is a capsule, it may further contain a liquid carrier such as oil.

A medicament containing, as an active ingredient, the compound represented by formula (I) of one embodiment of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, may be formulated as a depot preparation. can also In this case, the medicament of this aspect in the form of a depot formulation can be implanted, for example, subcutaneously or intramuscularly, or administered by intramuscular injection. By applying the medicament of this aspect to a depot preparation, the bone metabolism inhibitory activity of the compound represented by formula (I) of one aspect of the present invention can be sustained over a long period of time.

A medicament containing, as an active ingredient, the compound represented by the formula (I) of one embodiment of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, is one or more pharmaceutically useful It can also be used in combination with other drugs. In this case, the medicament of this aspect includes the compound represented by formula (I) of one aspect of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more It will be in the form of a combination drug containing the other drug. The concomitant drug is a compound represented by formula (I) of one embodiment of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more of the other drugs The compound represented by formula (I) of one embodiment of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof It may be in the form of a pharmaceutical composition for use in combination with one or more of the other drugs described above. When the medicament of this aspect is in the form of a combination medicament as described above, the compound represented by formula (I) of one aspect of the present invention or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvent thereof and one or more other drugs, which may be provided in the form of a single formulation, or a pharmaceutical combination comprising multiple formulations separately formulated with one or more other drugs, or It may be provided in the form of a kit. When in the form of a pharmaceutical combination or kit, each formulation can be administered simultaneously or separately (eg, sequentially).

The compound represented by formula (I) of one embodiment of the present invention has high BMP signaling inhibitory activity. The BMP signaling pathway mediated by the ALK2 receptor is considered to be not only the main cause of FOP, but also related to other diseases caused by abnormal bone metabolism. Therefore, a medicament containing, as an active ingredient, the compound represented by formula (I) of one embodiment of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof is Numerous conditions, diseases and/or disorders resulting from metabolic abnormalities can be prevented or treated. Said conditions, diseases and/or disorders include, but are not limited to, fibrodysplasia ossificans progressive (FOP), malignant hypercalcemia, dysostosis, abnormally increased bone turnover. Rotation, ectopic calcification, Ehlers-Danlos syndrome, Osgood-Schlatter disease, traumatic ectopic ossification, hallux valgus, jawbone facial bone reconstruction, articular cartilage calcification, rheumatoid arthritis, pseudomalignant displaced ossification, Kienbock disease, proximal phalanx, muscular contracture, rickets, glucocorticoid-induced osteoporosis, open tibia fracture, cervical spondylosis, myositis ossificans focalis, primary pulmonary hypertension, primary parathyroid function hypertension, posterior longitudinal ligament ossification, hyperthyroidism, bone lengthening, metaphyseal dysplasia, osteogenesis imperfecta, osteogenic mass, osteosclerosis, osteosclerotic metastatic bone tumor, posterior longitudinal ligament bone Metaplasia, fracture, osteoarthritis, osteoporosis, osteomalacia, osteochondrodysplasia, osteosarcoma, Paget's disease of bone, osteomalacia, osteomalacia, ossification due to insufficient oxygen supply, periodontal disease, teeth loss, juvenile polyposis, gastrointestinal cancer, microtia cartilage defect, ossification neurosis, myositis ossans progressive, fibrodysplasia ossificans progressive, renal osteodystrophy, spinal canal stenosis, Kyphosis, lordosis, scoliosis, spinal metaphyseal dysplasia, spinal epiphyseal dysplasia, spondylolisthesis, spondylolisthesis, congenital torticollis congenita, congenital hip dislocation, multiple congenital Arthrogryposis, congenital clubfoot, scoliosis, femoral head necrosis, osteopetrosis, multiple synostosis, multiple myeloma, brachydactyly, metacarpal shortening, metacarpal shortening, metacarpal shortening , intervertebral disc degeneration, metastatic bone disease, metastatic bone tumor, idiopathic femoral head necrosis, clubfoot, chondroectodermal dysplasia, achondroplasia, achondroplasia, Hurler syndrome, postmenopausal osteoporosis, Perthes disease , osteoarthritis, spondylosis osteoarthritis, degenerative spondylolisthesis, periprosthetic osteolysis, shortened distal phalanges, Marfan syndrome, rheumatoid arthritis, chronic renal failure, aneuria, Meller-Barrow disease, Morquio disease, lumbar spine Disc herniation and osteochondritis dissecans can be mentioned. Preferably, said condition, disease and/or disorder is FOP or traumatic heterotopic ossification. The symptoms, diseases or disorders can be prevented or treated by administering the medicament of this aspect to a subject in need of prevention or treatment of the symptoms, diseases or disorders.

A medicament containing, as an active ingredient, the compound represented by formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof according to one embodiment of the present invention is a bone It can be applied to various subjects in need of prevention or treatment of the aforementioned symptoms, diseases and/or disorders caused by metabolic disorders. The subject is a human or non-human mammal (e.g., warm-blooded animals such as pigs, dogs, cows, rats, mice, guinea pigs, rabbits, chickens, sheep, cats, monkeys, baboons, or chimpanzees) subjects or patients. Preferably. By administering the pharmaceutical of this aspect to the subject, various symptoms, diseases and/or disorders caused by abnormal bone metabolism in the subject can be prevented or treated.

As used herein, "prevention" means to substantially prevent the occurrence (development or manifestation) of symptoms, diseases and/or disorders. In addition, as used herein, “treatment” means suppressing (for example, suppressing progression), relieving, repairing and/or curing a symptom, disease and/or disorder that has occurred (onset or manifested).

The compound represented by formula (I) of one aspect of the present invention is a symptom, disease and/or disorder caused by abnormal bone metabolism involving BMP signaling described above (e.g., FOP or traumatic ectopic ossification) can be used to prevent or treat said condition, disease and/or disorder. Therefore, the medicament of this embodiment is preferably a medicament for use in the prevention or treatment of the symptoms, diseases and/or disorders described above, and is used in the prevention or treatment of FOP or traumatic heterotopic ossification. More preferably, it is a medicament for use. Represented by Formula (I) of one embodiment of the present invention, by using the pharmaceutical of this embodiment for the prevention or treatment of the symptoms, diseases and/or disorders caused by abnormal bone metabolism involving BMP signaling Such conditions, diseases and/or disorders can be prevented or treated through the BMP signaling inhibitory activity of the compounds.

The compound represented by formula (I) of one aspect of the present invention is used in subjects having the symptoms, diseases and/or disorders (e.g., FOP or traumatic heterotopic ossification) described above. and/or for the prevention or treatment of disorders. Therefore, another aspect of the present invention provides a subject in need of prevention or treatment of the symptoms, diseases and/or disorders described above, in an effective amount of a compound of formula (I) of the present invention or A method for preventing or treating the disease or condition, comprising administering a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. Preferably said condition, disease and/or disorder is FOP or traumatic heterotopic ossification. administering a compound represented by formula (I) of one embodiment of the present invention to a subject in need of prevention or treatment of the symptoms, diseases and/or disorders caused by abnormal bone metabolism involving BMP signaling; Accordingly, the symptoms, diseases and/or disorders can be prevented or treated via the BMP signaling inhibitory activity of the compound represented by formula (I) of one embodiment of the present invention.

Another aspect of the invention is the formula of one aspect of the invention for use in the prevention or treatment of the conditions, diseases and/or disorders described above (e.g. FOP or traumatic heterotopic ossification). A compound represented by (I), a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. Another aspect of the present invention is the use of the present invention for the manufacture of a medicament for use in the prevention or treatment of the conditions, diseases and/or disorders described above (e.g. FOP or traumatic heterotopic ossification). or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof. Preferably said condition, disease and/or disorder is FOP or traumatic heterotopic ossification. By using the compound represented by the formula (I) of one embodiment of the present invention for the prevention or treatment of the symptoms, diseases and/or disorders caused by abnormal bone metabolism involving BMP signaling, Said symptoms, diseases and/or disorders can be prevented or treated through the BMP signaling inhibitory activity of the compound represented by Formula (I) in one embodiment.

Administration of a medicament containing, as an active ingredient, the compound represented by formula (I) of one embodiment of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, to a subject, particularly a human patient If so, the precise dose and regimen (e.g., dose, frequency and/or route of administration) will depend on the subject's age, sex, and the precise state of the condition, disease and/or disorder to be prevented or treated (e.g., severe In view of many factors such as dosage, administration route, etc., the final decision should be made by the attending physician, taking into account the therapeutically effective dose, administration frequency, administration route, and the like. Therefore, in the medicament of this aspect, the active ingredient, the compound represented by formula (I), is administered to the subject in a therapeutically effective amount and at a frequency. For example, when the medicament of this embodiment is administered to a human patient, the dose of the compound represented by formula (I), which is an active ingredient, is usually in the range of 0.001 to 100 mg/kg body weight per administration. Yes, typically in the range of 0.01 to 10 mg/kg body weight per dose, especially in the range of 0.1 to 10 mg/kg body weight per dose. In addition, the administration frequency of the medicament of this aspect can be, for example, once or multiple times a day, or once every several days. In addition, the administration route of the medicament of this aspect is not particularly limited, and it may be administered orally or parenterally (e.g., rectal, transmucosal, intestinal, intramuscular, subcutaneous, intramedullary, intrathecal , directly intraventricularly, intravenously, intravitreally, intraperitoneally, intranasally or intraocularly) in single or multiple doses. By using the medicament of this aspect in the dosage and usage described above, bone metabolism involving BMP signaling is suppressed via the BMP signaling inhibitory activity of the compound represented by formula (I) of one aspect of the present invention. The symptoms, diseases and/or disorders resulting from the abnormality can be prevented or treated.

EXAMPLES The present invention will be described in more detail below using examples. However, the technical scope of the present invention is not limited to these examples.

<I. PRODUCTION OF NOVEL COMPOUNDS BY CULTURAL MEANS>
[Isolation of BF-0343 strain]
The BF-0343 strain is a filamentous fungus isolated from soil in Shizuoka Prefecture, Japan. Based on the morphological characteristics, culture properties and physiological properties of the BF-0343 strain, comparison with known strains was performed. As a result, this strain was found to be a novel filamentous fungus strain belonging to the genus Penicillium. This strain was named BF-0343 strain. This strain was registered as BF-0343 on November 28, 2018 at the Patent Microorganism Depositary Center, National Institute of Technology and Evaluation (2-5-8-122 Kazusa Kamatari, Kisarazu City, Chiba Prefecture 292-0818, Japan). room) (accession number NITE P-02833). The mycological properties of the BF-0343 strain are as follows.

1. Morphological features
The BF-0343 strain grew well on a medium such as a nutrient agar medium such as a PDA medium. Colonies grown on PDA medium at 27° C. under aerobic conditions for one week were observed under a microscope, and the surfaces of the colonies were velvety to powdery.

2. Culture properties
Colonies of strain BF-0343 grown on PDA medium at 27°C under aerobic conditions for one week have dark green (28F-4) to greenish white (28A-2) surfaces and gray undersurfaces. It was yellow (3B-5) to white (3A-1). The numbers in parentheses indicate color code numbers used in Kornerup Wanscher (Methuen handbook of colour, 3rd edition, London: Eyre Methuen, 1978). Also, no coloration of the medium due to the soluble pigment was observed.

3. Physiological properties
The optimum growth conditions for strain BF-0343 were aerobic conditions, pH 5.6±0.2, and temperature 25°C. In addition, the viable conditions for this strain were aerobic conditions, static culture, pH range of 3-9, temperature range of 25-27°C, and culture period of 1-3 weeks.

[Production Example I-1: Culture of BF-0343 strain]
One agar piece of BF-0343 strain cultured in PDA medium (3.9% potato dextrose agar (DIFCO), adjusted to pH 6.0) was suspended in 500 μL of sterile purified water. Inoculate the entire suspension into a 50 mL test tube containing 15 mL of PDB medium (2.4% potato dextrose broth (DIFCO), 0.1% agar (SSK Sales), adjusted to pH 6.0), It culture|cultivated using the shaking culture machine (140 rpm) at 27 degreeC for 3 days (seed culture). 100 mL of the same medium was dispensed into a 500 mL volume Erlenmeyer flask, and the resulting culture was inoculated at a concentration of 1%, and shake cultured using a rotary shaker (180 rpm) at 27°C for 7 days. (production culture).

[Production Example I-2: Purification of novel compound from culture medium of BF-0343 strain]
Ethanol (100 mL) was added to the culture solution (100 mL) obtained in Production Example I-1 and allowed to stand at 4°C for 24 hours. After that, the culture solution was filtered by suction to remove the cells, and the ethanol extract was recovered. The obtained ethanol extract was concentrated under reduced pressure to distill off the ethanol. The resulting water residue was extracted with ethyl acetate (200 mL). The ethyl acetate layer was collected and dried _{over Na2SO4} . After that, the ethyl acetate layer was dried under reduced pressure to obtain an ethyl acetate extract (61.1 mg). This extract was dissolved in a small amount of methanol and final purified using preparative high performance liquid chromatography (HPLC) (column: PEGASIL ODS SP100, 20φ×250 mm, Senshu Kagaku). For purification using HPLC, 20% aqueous acetonitrile was used as the mobile phase and the absorbance at UV 210 nm was monitored at a flow rate of 6 mL/min. A peak indicating activity was observed at a retention time of 22 minutes. After fractionating this peak and distilling off the solvent, the fraction was freeze-dried to isolate a pale yellow oily compound BF-0343 with a yield of 7.5 mg. The chemical structure of compound BF-0343 (5-(1-ethoxypropan-2-yl)-5-hydroxy-4-methoxyfuran-2(5H)-one) is shown below.

[Compound BF-0343]
₍ 1) _Properties : Pale yellow oil (2) Molecular formula: _C10H16O5
HREI-MS (m/z) [M] ⁺ calculated value 216.0998, measured value 216.0990
(3) Molecular weight: 216
Observed EI-MS (m/z) [M] ⁺ 216 (4) Ultraviolet absorption spectrum: measured in methanol solution:
λ _max (MeOH, ε): Absorption at 224 nm (7614) (5) Infrared absorption spectrum: Measured by the potassium bromide tablet method:
Characteristic absorption maxima at ν _max 3425, 2924, 1746, 1646, 1430, 1069, 919, 470 cm ^-1 etc. (6) Specific rotation: [α] _D ²⁵ -2.1° (c=0.1, methanol)
(7) Solubility in solvents: Soluble in methanol, chloroform, DMSO, etc. (8) Proton and carbon nuclear magnetic resonance spectra: chemical shifts of hydrogen measured in heavy chloroform with a 400 MHz nuclear magnetic resonance spectrometer manufactured by Agilent. (ppm) and carbon chemical shifts (ppm) are given below:
_δH : 0.71 (3H), 1.23 (3H), 2.50 (1H), 3.50-3.65 (3H), 3.82 (1H),
3.91 (3H), 5.06 (1H), 7.01 (1H) ppm
_δC : 11.2, 14.9, 36.5, 59.5, 67.3, 72.9, 89.7, 105.7, 170.4, 177.8 ppm.

<II. Pharmacological test of novel compounds>
[Test II-1: Alkaline phosphatase inhibitory activity test in C2C12 cell-derived osteoblasts]
C2C12 cells are a myoblast cell line established from the muscle regeneration part of adult mice. When BMP is applied to C2C12 cells, differentiation into myotubes is suppressed and differentiation into osteoblasts, ie, bone metabolism is promoted. Osteoblasts differentiated from C2C12 cells express alkaline phosphatase (T. Katagiri et al., J. Cell Biol., 127, 1755-1766, 1994). Here, when a specific compound is added to the differentiation induction system from C2C12 cells to osteoblasts induced by BMP, differentiation to osteoblasts, that is, bone metabolism is inhibited, and the expression level of alkaline phosphatase is decreased. Therefore, using alkaline phosphatase activity as an index, BMP signal transduction inhibitory activity against the BMP-induced differentiation induction system from C2C12 cells to osteoblasts, that is, bone metabolism inhibitory activity can be evaluated. The alkaline phosphatase inhibitory activity of the compounds prepared by the above procedure was investigated by the following procedure (T. Fukuda et al., J. Biol. Chem., 2009, 284, pp. 7149-7156).

Mouse muscle-derived C2C12 cells (C2C12 (R206H) cells, stored at Kitasato University, Faculty of Pharmaceutical Sciences, Department of Microbial Pharmaceuticals Manufacturing) stably transfected with a gene in which the 206th arginine of ALK2 is mutated to histidine were incubated with 15% fetal bovine serum ( The cells were suspended in Dulbecco's Modified Eagle's Medium (DMEM, Nacalai Tesque) containing FBS, Hyclone) and 1% penicillin/streptomycin (Invitrogen) to prepare a cell suspension of 7.5×10 ⁴ cells/mL. 100 μL of this cell suspension was seeded in each well of a 96-well microplate (0.75×10 ⁴ cells/100 μL/well). After seeding the cells, the microplate was transferred to a carbon dioxide gas incubator, and the cells were cultured for 24 hours under the conditions of 37°C and 5.0% CO ₂ . The medium in each well was replaced with the above medium (100 μL) containing 10 ng/mL recombinant human BMP-4 (R & D Systems). Test compounds at given final concentrations (462-4.62 μM) were added as 1.0 μL methanol solutions to given wells. Cells were then cultured for 48 hours at 37° C. and 5.0% CO ₂ . After the culture was completed, the culture supernatant was removed. Cells were washed once with phosphate buffered saline (PBS, pH 7.3). 100 μL of a mixed solution of acetone:ethanol=1:1 was dispensed into each well and allowed to stand for 1 minute to fix the cells. Cells were washed three times with PBS (pH 7.3). Dissolve 1 tablet (5 mg) of 4-nitrophenyl phosphate disodium salt hexahydrate (SIGMA) in 5 mL of reaction buffer (0.1 M diethanolamine-HCl and 0.5 mM MgCl ₂ , pH 10.0). to prepare a 1 mg/mL 4-nitrophenyl phosphate (4-NPP) solution. 100 μL of 1 mg/mL 4-NPP solution was dispensed into each well. The microplate was shaken at room temperature for 1 hour using a plate mixer (MICROMIXER MX-5, Sanko Junyaku Co., Ltd.). After that, 50 μL of 3 M sodium hydroxide aqueous solution was added to each well. The microplate was shaken for 5 minutes at room temperature. The absorbance of the solution in each well at a wavelength of 405 nm was measured using a microplate reader (Power Wave x340, Bio-Tek Instruments). The absorbance of wells tested in the same manner as above except that the same amount of methanol was added instead of the test compound was used as the absorbance of the control group. The absorbance of the wells tested in the same manner as above except that the cells were not seeded was used as the background value. The BMP signaling inhibition rate of each test compound was calculated based on the following formula. A dose-response curve for alkaline phosphatase (ALP) inhibition was generated for each test compound and the 50% inhibition concentration (ALP-IC ₅₀ value) for each test compound was calculated.

[Test II-2: Cytotoxicity test in C2C12 cell-derived osteoblasts]
Cytotoxicity of compounds prepared by the above procedure was evaluated by the MTT assay method (Mosmann et al., J Immunol Methods, 1983, 65:55-63) using thiazolyl blue tetrazolyl bromide (MTT). C2C12(R206H) cells were suspended in DMEM medium containing 15% FBS and 1% penicillin/streptomycin to prepare a cell suspension of 7.5×10 ⁴ cells/mL. 100 μL of this cell suspension was seeded in each well of a 96-well microplate (0.75×10 ⁴ cells/100 μL/well). After seeding the cells, the microplate was transferred to a carbon dioxide gas incubator, and the cells were cultured for 24 hours under the conditions of 37°C and 5.0% CO ₂ . The medium in each well was replaced with 100 μL of the above medium containing 10 ng/mL recombinant human BMP-4. To given wells, given final concentrations of test compounds were added as a 1.0 μL methanol solution. Cells were then cultured for 2 days at 37°C and 5.0% _CO2 . After the culture was completed, the culture supernatant was removed. Cells were washed once with PBS. To each well was added 10 μL of 5.5 mg/mL MTT reaction solution (PBS containing 5.5 mg/mL MTT, pH 7.3). Cells were cultured for 3 hours at 37° C., 5.0% CO ₂ . To each well was added 90 μL of lysate (40% N,N,-dimethylformamide, 2% acetic acid, 20% sodium dodecyl sulfate, and 0.03 N hydrochloric acid). The microplate was shaken for 3 hours at room temperature using a plate mixer. The absorbance of the solution in each well at a wavelength of 570 nm was measured using a microplate reader. The absorbance of wells tested in the same manner as above except that the same amount of methanol was added instead of the test compound was used as the absorbance of the control group. The absorbance of the wells tested in the same manner as above except that the cells were not seeded was used as the background value. Cell viability in the presence of each test compound was calculated based on the following formula. For each test compound, a dose-response curve for viability in the MTT assay was constructed and the 50% viability concentration (MTT- _IC50 value) for each test compound was calculated.

[Results of test II-1 and II-2]
The ALP-IC ₅₀ value for compound BF-0343 was 19.8 μM. Also, the survival rate of compound BF-0343 in the MTT assay was 69% at a concentration of 462 μM, and the MTT-IC ₅₀ value was >462 μM. The above results demonstrate that compound BF-0343 exhibits substantially no cytotoxicity at concentrations sufficient to inhibit differentiation of osteoblasts.

Compounds FKI-5513-1 and FKI-5513-2 described in Patent Document 1 (Patent No. 5725469) showed ALP-IC ₅₀ values of 83 and 187 μM in a test similar to Test II-1. It has been known. Therefore, it was revealed that the compound BF-0343 of the present invention has a bone metabolism inhibitory activity that greatly exceeds that of the compound described in Patent Document 1.

It should be noted that the present invention is not limited to the above-described embodiments, and includes various modifications. For example, the above-described embodiments have been described in detail for easy understanding of the present invention, and are not necessarily limited to those having all the described configurations. Moreover, it is possible to add, delete and/or replace part of the configuration of each embodiment with another configuration.

Claims (10)

Formula (I):

[In the formula,
^R1 is ethoxyl ,
^R2 is hydrogen,
^R3 is methyl ,
^R4 is methyl . ]
A compound represented by or a salt thereof, or a solvate thereof. A method for producing the compound according to claim 1 , a salt thereof, or a solvate thereof,
Penicillium strain BF-0343 (Accession No. NITE P-02833 ) having the ability to produce the compound represented by formula (I) is cultured in a medium to produce the compound represented by formula (I) a compound accumulation step of accumulating in said medium;
A compound purification step of purifying the compound represented by formula (I) obtained in the compound accumulation step from the culture of the microorganism;
The above method, comprising A microorganism which is Penicillium strain BF-0343 (accession number NITE P-02833 ), which has the ability to produce the compound represented by formula (I) according to claim 1 , a salt thereof, or a solvate thereof. A bone metabolism inhibitor comprising the compound according to claim 1 , a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient. 10. A medicine comprising the compound according to claim 1 , a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof as an active ingredient. 6. The medicament according to claim 5 , for use in preventing or treating one or more symptoms, diseases or disorders caused by abnormal bone metabolism. 7. The medicament according to claim 6 , wherein the one or more symptoms, diseases or disorders caused by abnormal bone metabolism are fibrodysplasia ossificans progressive (FOP) or traumatic heterotopic ossification. A pharmaceutical composition comprising the compound of claim 1 or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable solvate thereof, and one or more pharmaceutically acceptable carriers. 9. The pharmaceutical composition according to claim 8 , for use in preventing or treating one or more symptoms, diseases or disorders caused by abnormal bone metabolism. 10. The pharmaceutical composition according to claim 9 , wherein the one or more symptoms, diseases or disorders caused by abnormal bone metabolism is fibrodysplasia ossificans progressive (FOP) or traumatic heterotopic ossification.