JP2014101274A - Pharmaceutical composition using mimic peptide of tnf receptor loop peptide comprising new crosslinked structure - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a new mimic peptide of TNF receptor loop peptide (WP9QY peptide) having improved properties, more specifically to provide an osteoporosis therapeutic drug comprising a WP9QY peptide mimic having more desirable properties.SOLUTION: A mimic peptide of TNF receptor loop peptide having a new crosslinked structure with -NR- in a crosslinked structure has osteoclast-differentiation inhibitory activity. In addition, an osteoclast-proliferation inhibitor using the peptide and a pharmaceutical composition comprising the peptide are provided.

Description

  The present invention relates to an osteoclast growth inhibitor using a mimetic peptide of a TNF receptor loop peptide containing a novel cross-linked structure. The present invention also relates to a pharmaceutical composition using the peptide.

Almost all physiological processes are based on molecular recognition such as peptides or proteins and other biologically active ingredients. To date, many proteins with important biological functions such as hormones, enzymes, inhibitors, enzyme substrates, neurotransmitters, immunomodulators have been discovered. However, since the protein has a large molecular weight, it is difficult not only to synthesize it but also to be used as a pharmaceutical product. Therefore, elucidation of its function at the molecular level has been made.
The action of a protein in a living body is generally activated via a receptor. Receptors are also proteins and the binding or contact of both (ligand and receptor) is recognized as important for physiological effects. Therefore, the site involved in the binding between the ligand and the receptor is identified, a peptide mimicking that part is synthesized, the function of the protein is analyzed, and the peptide as a blocking substance that inhibits the action of the protein or the protein Has attracted attention.

However, it is known that the portion involved in the binding or contact between the ligand / receptor takes a configuration in vivo, and the conformation of the binding or contact portion is important. For this reason, it has been attempted to create a peptide having a conformation in a portion involved in binding or contact.
Overall control in peptide conformation is possible by limiting the flexibility of the peptide chain by cyclization. Cyclization of a biologically active peptide not only improves its metabolic stability and selectivity for the receptor, but also allows conformational analysis of the peptide by making it a homogeneous conformation. The form of cyclization is the same as found in natural cyclic peptides. For example, side chain-side chain cyclization or side chain-end group cyclization can be mentioned. For cyclization, amino acid side chains that are not involved in ligand / receptor recognition can be linked to each other or to the peptide backbone. In another embodiment, terminal-terminal cyclization can be mentioned. In such a case, a completely cyclic peptide is obtained.

  Cross-linking techniques are indispensable for these cyclizations. Representative examples of cyclization include crosslinking via disulfide bonds (SS bonds), amide bonds, thioether bonds, and olefin bonds. A more specific example is the cyclization of two penicillamine residues linked by a disulfide bridge (Mosberg et al., PNAS US, 80: 5871, 1983), where an amide bond is formed between lysine and aspartic acid. A cyclization by formation (Flora et al., Bioorg. Med. Chem. Lett. 15 (2005) 1065-1068) or an amino acid derivative containing a cross-linked site previously introduced with a thioether bond is introduced into the peptide bond, Cyclized by the condensation reaction of (Melin et al. US 6,143,722) and (S) -α-2′-pentenyl) alanine introduced into the main chain using an olefin metathesis reaction to cyclize by crosslinking alanines ( Schafmeister et al., J. Am. Chem. Soc., 122, 5891-5892, 2000). In addition, as a method that does not require modification of the side chain of the peptide in order to control conformation, a crosslinked structure starting from nitrogen constituting the amide of the peptide main chain skeleton is also known. (Gilon et al. Bioplymers 31: 745, 1991).

  Peptides with controlled conformation can be expected for many pharmacological uses. However, there are the following problems in the development of peptides as pharmaceuticals. That is, under physiological conditions, many of the peptides are degraded by specific and non-specific peptidases, resulting in poor metabolic stability, and b) because of their relatively high molecular weight, Poor absorption, c) fast excretion through the liver and kidney, and d) untargeted tissue because the peptide is structurally flexible and receptors for the peptide can be widely distributed in organisms -Undesirable side effects occur in the organ.

Receptor Activator of NF-κB (also known as RANK, TRANCER or TNFRSF11A (tumor necrosis factor receptor superfamily member 11a)) belonging to the TNF receptor superfamily includes osteoclasts, dendritic cells, mammary epithelial cells, etc. It is expressed in. RANK ligands (RANKL, or OPGL, ODF, TRANCE, and TNFSF11 (also known as TNF ligand superfamily member 11)) are TNF-like proteins and are expressed in osteoblasts, bone interstitial cells, and T cells. Has been. RNAK and RANKL have also been reported to be required for osteoclast differentiation, dendritic cell differentiation and mammary gland development in vivo and in vitro. TNFα, which belongs to the TNF family, not only works directly on bone marrow cells to promote osteoclast formation, but also works on osteoblasts to increase the level of RANKL and indirectly amplify the osteoclast-forming response of bone marrow cells. And greatly contribute to bone loss induced by ovariectomy and inflammation. RANKL also increases bone resorption by osteoclasts.
The WP9QY peptide is a cross-linked peptide having a peptide sequence consisting of 5 amino acids in its ring, which resembles the sequence structure of the functional sites of TNF receptor and RANK, and has the following structure (Japanese Patent Publication No. 2003-505503, And Special Table 2003-505514).

ＷＰ９ＱＹペプチドは、ＴＮＦαにより誘導される細胞毒性を阻害することが報告されている（高橋ら、Nat. Biotechnol. 1997, 15: 1266-1270）。ＷＰ９ＱＹペプチドはまた、ＲＡＮＫＬによって誘導されるシグナルをブロックし、骨吸収および骨損失を阻害することが報告されている（青木ら、J. Clin Invest. 2006; 116(6):1525-1534）。また、炎症性骨吸収を引き起こすリウマチ間接モデルにおいてＷＰ９ＱＹペプチドが同用量の抗ＴＮＦ抗体投与と比べて骨吸収抑制効果が優れていることが報告されている（斉藤ら、Rheumatoid Arthritis, Vol.56, No.4, 1164-1174）。さらに、歯周病原因細菌による炎症性骨破壊をＷＰ９ＱＹペプチド投与により抑制することも報告されている（鈴木ら、J. Periodontal Res., 2006 Apr. 41(2):81-91）。

WP9QY peptide has been reported to inhibit cytotoxicity induced by TNFα (Takahashi et al., Nat. Biotechnol. 1997, 15: 1266-1270). The WP9QY peptide has also been reported to block RANKL-induced signals and inhibit bone resorption and bone loss (Aoki et al., J. Clin Invest. 2006; 116 (6): 1525-1534). In addition, it has been reported that the WP9QY peptide is superior to the administration of the same dose of anti-TNF antibody in the rheumatoid indirect model causing inflammatory bone resorption (Saito et al., Rheumatoid Arthritis, Vol. 56, No. 4, 1164-1174). Furthermore, it has also been reported that inflammatory bone destruction caused by periodontal disease-causing bacteria is suppressed by administration of WP9QY peptide (Suzuki et al., J. Periodontal Res., 2006 Apr. 41 (2): 81-91).

  As used herein, a “peptidomimetic” is a compound that can, as a receptor ligand, mimic (agonist) or mask (antagonist) the biological effects of a peptide at the receptor level. To obtain a peptidomimetic as the most likely agonist, a) metabolic stability, b) good bioavailability, c) high receptor affinity and receptor selectivity, and d) Factors such as minimal side effects need to be considered. Also, from a pharmacological and medical point of view, it is often desirable not only to mimic the effects of peptides at the receptor level (acting action), but also to mask the receptor (antagonism) if necessary. The same pharmacological considerations that should be considered for designing a peptidomimetic as an agonist above can also be applied to the design of peptide antagonists.

Special table 2003-505503 gazette Special table 2003-505514 gazette

Mosberg et al., P.N.A.S.US, 80: 5871, 1983 Charpentier et al., J. Med. Chem. 32: 1184, 1989 Rodriguez et al., Int. J. Pept. Protein Res. 35: 441, 1990 Schafmeister et al., J. Am. Chem. Soc., 122, 5891-5892, 2000 Goodman et al., J. Org. Chem. 2002, 67, 8820-8826 Takahashi et al., Nat. Biotechnol. 1997, 15: 1266-1270 Aoki et al., J. Clin Invest. 2006; 116 (6): 1525-1534 Saito et al., Rheumatoid Arthritis, Vol.56, No.4, 1164-1174 Suzuki et al., J. Periodontal Res., 2006 Apr. 41 (2): 81-91

.
WP9QY peptide is known as a peptidomimetic that inhibits cytotoxicity induced by TNFα and inhibits bone resorption, but there is a problem that disulfide bonds are generally cleaved by reductase present in the living body. Further improved mimetic peptides were desired. Although other cyclization forms exist, those having an amide in the cross-linking have a problem that they are cleaved by an enzyme that cleaves the amide structure existing in the living body. Furthermore, free design of the crosslink and free change to the crosslink are desired to improve the physiological activity of the peptide, but there is a problem that it is difficult in the conventional cyclization format.
An object of the present invention is to provide an osteoporosis therapeutic agent comprising a WP9QY peptidomimetic having more desirable properties.

As a result of intensive studies to solve the above problems, the present inventors synthesized a WP9QY peptide mimic having a novel cross-linked structure therein, and that it has an osteoclast differentiation inhibitory activity. Confirmed and completed the present invention.
That is, the present invention provides an osteoclast growth inhibitor using the novel WP9QY peptide mimic described below and a pharmaceutical composition containing the peptide.
1. Osteoclast growth inhibitor comprising a peptide represented by the following chemical formula

（ここで、Ｚ₁およびＺ₃は、それぞれ独立に、水素原子、無置換または置換された炭素数１〜８のアシル基、無置換または置換された炭素数１〜８のアルキル基および−Ｃ（＝Ｏ）−ＣＨ₂ＣＨ₂（ＯＣＨ₂ＣＨ₂）_nＯＣＨ₂ＣＨ₂ＯＣＨ₃で表される分子量１００〜１０，０００Ｄａのポリエチレングリコールからなる群より選ばれ、Ｚ₂はヒドロキシル基、アミノ基、無置換または置換された炭素数１〜８のモノアルキルアミノ基、−ＮＨ−ＣＨ₂ＣＨ₂（ＯＣＨ₂ＣＨ₂）_nＯＣＨ₂ＣＨ₂ＯＣＨ₃で表される分子量１００〜１０，０００Ｄａのポリエチレングリコールからなる群より選ばれる）
２．下記の化学式で表されるペプチドからなる群より選ばれる上記１に記載の破骨細胞の分化または増殖阻害剤、

Wherein Z ₁ and Z ₃ are each independently a hydrogen atom, an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and —C (═O) —CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ is selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da and Z ₂ is a hydroxyl group or an amino group , An unsubstituted or substituted monoalkylamino group having 1 to 8 carbon atoms, and a polyethylene having a molecular weight of 100 to 10,000 Da represented by —NH—CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ Selected from the group consisting of glycols)
2. The osteoclast differentiation or proliferation inhibitor according to the above 1, which is selected from the group consisting of peptides represented by the following chemical formula:

および

and

（ここで、Ａｃはアセチル基を表し、ポリエチレングリコールは、５００〜２０００Ｄａの数平均分子量をもつポリエチレングリコールを表す）。

(Here, Ac represents an acetyl group, and polyethylene glycol represents polyethylene glycol having a number average molecular weight of 500 to 2000 Da).

3. 3. A pharmaceutical composition comprising as an active ingredient at least one of the peptides represented by the chemical formula according to 1 or 2 above.
4). It is a therapeutic agent for diseases selected from the group consisting of osteoporosis, Paget's disease, rheumatoid arthritis and other forms of inflammatory arthritis, osteoarthritis, prosthetic breakage, osteolytic tumors, myeloma and tumor metastasis to bone The pharmaceutical composition according to 3 above.
5. A method of treating a disease selected from the group consisting of osteoporosis, Paget's disease, rheumatoid arthritis and other forms of inflammatory arthritis, osteoarthritis, prosthetic breakage, osteolytic tumors, myeloma and tumor metastasis to bone. A method of treatment comprising administering to a mammal in need of such treatment at least one of the chemical formulas described in 1 or 2 in a therapeutically effective amount.

  The present invention provides a pharmaceutical composition comprising a WP9QY peptidomimetic with improved properties, eg, increased resistance to peptidases or increased inhibitory activity against osteoclasts.

FIG. 1 schematically shows an example of synthesis of a WP9QY peptide mimetic of the present invention. FIG. 2 shows schematically a synthesis example of the WP9QY peptidomimetic of the present invention. FIG. 3 shows a synthesis example of a WP9QY peptide mimetic having a thioether bridge as a reference example. FIG. 4 shows a synthesis example of a WP9QY peptide mimetic having an olefinic bridge as a reference example. FIG. 5 shows the osteoclast differentiation-inhibiting activity of the WP9QY peptide mimics (Bdev-2, Bdev-5, Bdev-6, Bdev-8) of the present invention. FIG. 6 shows the osteoclast differentiation inhibitory activity of the WP9QY peptide mimics (Bdev-2, Bdev-3) of the present invention.

  The WP9QY peptidomimetic of the present invention is a cyclic peptide, and has the same amino acid sequence as that of the WP9QY peptide in the ring, and has a —NR— bond in the crosslinked structure. Specifically, the WP9QY peptidomimetic of the present invention is a peptide having the following structure.

（ここで、Ｚ₁およびＺ₃は、それぞれ独立に、水素原子、無置換または置換された炭素数１〜８のアシル基、無置換または置換された炭素数１〜８のアルキル基、および−Ｃ（＝Ｏ）−ＣＨ₂ＣＨ₂（ＯＣＨ₂ＣＨ₂）_nＯＣＨ₂ＣＨ₂ＯＣＨ₃で表される分子量１００〜１０，０００Ｄａのポリエチレングリコールからなる群より選ばれ、Ｚ₂はヒドロキシル基、アミノ基、無置換または置換された炭素数１〜８のモノアルキルアミノ基、−ＮＨ−ＣＨ₂ＣＨ₂（ＯＣＨ₂ＣＨ₂）_nＯＣＨ₂ＣＨ₂ＯＣＨ₃で表される分子量１００〜１０，０００Ｄａのポリエチレングリコールからなる群より選ばれる）

(Wherein Z ₁ and Z ₃ are each independently a hydrogen atom, an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and- C (═O) —CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ is selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da and Z ₂ is a hydroxyl group, an amino group Group, an unsubstituted or substituted monoalkylamino group having 1 to 8 carbon atoms, a molecular weight of 100 to 10,000 Da represented by —NH—CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ (Selected from the group consisting of polyethylene glycol)

Z ₁ and Z ₃ are each independently a hydrogen atom, an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and —C (═O). It is selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da represented by —CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ , preferably a hydrogen atom, substituted or substituted An acyl group having 1 to 6 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 6 carbon atoms, or —C (═O) —CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ Polyethylene glycol having a molecular weight of 300 to 5,000 Da, more preferably a hydrogen atom, a substituted or substituted acyl group having 1 to 4 carbon atoms, an unsubstituted or substituted carbon atom having 1 to 4 carbon atoms. Kill group, or is C (= O) polyethylene glycol having a molecular weight 500~2,000Da represented by _{-CH 2 CH 2 (OCH 2 CH} 2) n OCH 2 CH 2 OCH 3.
Z ₂ is represented by a hydroxyl group, an amino group, an unsubstituted or substituted monoalkylamino group having 1 to 8 carbon atoms, —NH—CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH _3. Selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da, preferably a hydroxyl group, an amino group, an unsubstituted or substituted monoalkylamino group having 1 to 6 carbon atoms, —NH—CH ₂ CH ₂ (OCH ₂ CH ₂ ) _n Polyethylene glycol having a molecular weight of 300 to 5,000 Da represented by OCH ₂ CH ₂ OCH ₃ , more preferably a hydroxyl group, an amino group, an unsubstituted or substituted carbon number of 1 to 4 monoalkylamino group, molecular weight 500～2,00 represented by _{-NH-CH 2 CH 2 (OCH} 2 CH 2) n OCH 2 CH 2 OCH 3 Polyethylene glycol Da.
One of Z ₁ and Z ₃ is an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and —C (═O) —CH. It is preferably selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da represented by ₂ CH ₂ (OCH ₂ CH ₂ ) _n OCH ₂ CH ₂ OCH ₃ , and the other is selected from a hydrogen atom or an acetyl group. .
Further, Z ₁ is hydrogen, an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, or an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and Z ₂ is a hydroxyl group, an amino group, In the case of an unsubstituted or substituted acyl group having 1 to 8 carbon atoms or an unsubstituted or substituted monoalkylamino group having 1 to 8 carbon atoms, Z ₃ represents —C (═O) —CH ₂ CH ₂ ( OCH ₂ CH ₂₎ is preferably a polyethylene glycol having a molecular weight 100~10,000Da represented by _n OCH ₂ CH ₂ OCH _3.

In the present specification, “inhibition of osteoclast proliferation” used when expressing the action of the WP9QY peptidomimetic of the present invention refers to immature osteoclasts that are mature cells when the growth or proliferation of osteoclasts is inhibited. It is used to include any of the cases where it is inhibited from differentiating.
The WP9QY peptidomimetic of the present invention is effective for diseases involving TNFα / TNF receptors and also useful for diseases involving RANK / RANKL. Diseases can include, for example, osteoporosis, Paget's disease, rheumatoid arthritis and other forms of inflammatory arthritis, osteoarthritis, prosthetic damage, osteolytic tumors, myeloma and tumor metastasis to bone, Without limitation, diseases involving TNFα / TNF receptor and / or RANK / RANKL are also included.

The peptide of the present invention may be a pharmaceutical composition containing any known pharmaceutically acceptable carrier.
The peptide of the present invention may be in the form of a pharmaceutically acceptable salt.
The peptides of the present invention can be administered orally or parenterally. In the case of oral administration, it can be administered in the form of capsules, tablets, granules, liquids and the like. In the case of parenteral administration, it is administered in the form of injection solution, infusion preparation or the like.
The peptide of the present invention can be used alone or in combination with other osteoporosis therapeutic / prophylactic agents such as organic bisphosphonates.

The peptides of the present invention can be administered to, for example, humans or other warm-blooded animals (eg, rats, mice, rabbits, dogs, monkeys, etc.). Appropriate doses of the peptides of the present invention will vary depending on factors such as the disease or disorder being treated, the route of administration, the age and weight of the patient being treated. In the case of oral administration, the active ingredient is generally about 0.01 mg to 1000 mg, preferably about 1.0 to 200 mg per day, usually in human adults. About 0.01 to 100 mg, preferably about 0.1 to 10 mg of the active ingredient is administered by intravenous injection.
The preparation of the present invention is preferably administered parenterally, for example, intramuscularly, intraperitoneally, intravenously, subcutaneously, and the like.
EXAMPLES Hereinafter, although an Example demonstrates this invention in detail, this invention is not limited to these.

  The following abbreviations were used in the following examples.

Synthesis of WP9QY Peptidomimetic A bridge that is a WP9QY peptidomimetic with — (CH ₂ ) ₄ —NH— (CH ₂ ) ₄ — as the bridging structure instead of a disulfide bond according to the synthetic schemes of FIGS. A peptide (Bdev7) was synthesized.
Hereinafter, each process will be described in detail.
Example 1 Synthesis of Synthetic Intermediate (Compound 14)
1-1. Synthesis of compound 13 as raw material Synthesis of compound 1

2,4-ジドコシロキシベンジルアルコール（「ＫｂＯＨ」と表記する） (7.69 g, 10.1 mmol) をCH₂Cl₂ (200 mL) に溶解し、Fmoc-Gly-OH (4.52 g, 15.2 mmol, 1.5 equiv)、DIPCI (3147 μＬ, 20.2 mmol, 2.0 equiv)、およびDMAP (12.3 mg, 0.101 mmol, 0.01 equiv) を加えて室温で30分間攪拌した。沈殿物をろ過し、ろ液を減圧下留去した。残渣にMeOHを加えて析出した沈殿物をろ過し、MeOH懸洗を2回 、CH₃CN懸洗を行い化合物1 (10.1 g, 96.7%) を得た。
化合物2の合成

2,4-didocosyloxybenzyl alcohol (referred to as “KbOH”) (7.69 g, 10.1 mmol) was dissolved in CH ₂ Cl ₂ (200 mL) and Fmoc-Gly-OH (4.52 g, 15.2 mmol, 1.5 equiv), DIPCI (3147 μL, 20.2 mmol, 2.0 equiv), and DMAP (12.3 mg, 0.101 mmol, 0.01 equiv) were added and stirred at room temperature for 30 minutes. The precipitate was filtered and the filtrate was distilled off under reduced pressure. MeOH was added to the residue, the deposited precipitate was filtered, MeOH rinsing twice and CH ₃ CN rinsing to obtain Compound 1 (10.1 g, 96.7%).
Synthesis of compound 2

化合物1 (10.1 g, 9.74 mmol) をTHF (200 mL) に溶解し、piperidne (1863 μＬ, 17.5 mmol, 1.8 equiv) およびDBU (1863 μＬ, 17.5 mmol, 1.8 equiv) を加えて室温で5分間攪拌した。反応液のpHが6前後になるまで濃塩酸を加えて減圧下溶媒を留去した。残渣にCH₃CNを加えて析出した沈殿物をろ過し、CH₃CN懸洗を2回行い化合物2 (8.21 g, 98.7%) を得た。
化合物3の合成

Compound 1 (10.1 g, 9.74 mmol) is dissolved in THF (200 mL), piperidne (1863 μL, 17.5 mmol, 1.8 equiv) and DBU (1863 μL, 17.5 mmol, 1.8 equiv) are added and stirred at room temperature for 5 minutes. did. Concentrated hydrochloric acid was added until the pH of the reaction solution reached around 6, and the solvent was distilled off under reduced pressure. CH ₃ CN was added to the residue, the deposited precipitate was filtered, and CH ₃ CN was washed twice to obtain Compound 2 (8.21 g, 98.7%).
Synthesis of compound 3

化合物2 (7.60 g, 8.96 mmol) をCH₂Cl₂ (180 mL) に溶解し、DIPEA (3589 μＬ, 20.6 mmol, 2.3 equiv) およびNsCl (2.59 g, 11.7 mmol, 1.3 equiv) を加えて室温で攪拌した。1時間後DIPEA (359 μＬ, 2.06 mmol, 0.2 equiv) を追加し室温で19分間攪拌した。反応液にＭｅＯＨ (30 mL) を加えた後、減圧下溶媒を留去した。残渣にCH₃CNを加えて析出した沈殿物をろ過し、CH₃CN懸洗を2回行い化合物3 (8.95 g, 99.9%) を得た。
化合物4の合成

Compound 2 (7.60 g, 8.96 mmol) was dissolved in CH ₂ Cl ₂ (180 mL), and DIPEA (3589 μL, 20.6 mmol, 2.3 equiv) and NsCl (2.59 g, 11.7 mmol, 1.3 equiv) were added at room temperature. Stir. After 1 hour, DIPEA (359 μL, 2.06 mmol, 0.2 equiv) was added, and the mixture was stirred at room temperature for 19 minutes. After adding MeOH (30 mL) to the reaction solution, the solvent was distilled off under reduced pressure. CH ₃ CN was added to the residue, and the deposited precipitate was filtered, and CH ₃ CN was washed twice to obtain Compound 3 (8.95 g, 99.9%).
Synthesis of compound 4

化合物3 (4.70 g, 4.71 mmol) および化合物A (3.11 g, 9.46 mmol, 2.0 equiv) をTHF (94 mL) に溶解し、PPh₃ (2.49 g, 9.50 mmol, 2.0 equiv) およびDEAD (4272 μＬ, 9.42 mmol, 2.0 equiv) を加えて室温で3時間攪拌した。PPh₃ (248 mg, 0.946 mmol, 0.2 equiv) およびDEAD (427 μＬ, 0.942 mmol, 0.2 equiv) を加えて1時間攪拌した。PPh₃ (245 mg, 0.934 mmol, 0.2 equiv) およびDEAD (427 μＬ, 0.942 mmol, 0.2 equiv) を加えて1時間15分攪拌した後、減圧下溶媒を留去した。残渣にCH₃CNを加えて析出した沈殿物をろ過し、CH₃CN懸洗を2回行い化合物4 (6.12 g, 99.2%) を得た。
化合物5の合成

Compound 3 (4.70 g, 4.71 mmol) and Compound A (3.11 g, 9.46 mmol, 2.0 equiv) were dissolved in THF (94 mL), and PPh ₃ (2.49 g, 9.50 mmol, 2.0 equiv) and DEAD (4272 μL, 9.42 mmol, 2.0 equiv) was added and stirred at room temperature for 3 hours. PPh ₃ (248 mg, 0.946 mmol, 0.2 equiv) and DEAD (427 μL, 0.942 mmol, 0.2 equiv) were added and stirred for 1 hour. PPh ₃ (245 mg, 0.934 mmol, 0.2 equiv) and DEAD (427 μL, 0.942 mmol, 0.2 equiv) were added and stirred for 1 hour and 15 minutes, and then the solvent was distilled off under reduced pressure. CH ₃ CN was added to the residue and the deposited precipitate was filtered, and CH ₃ CN was washed twice to obtain Compound 4 (6.12 g, 99.2%).
Synthesis of compound 5

化合物4 (5.65 g, 4.31 mmol) をTHF (40 mL) に溶解し、PhSH (1328 μＬ, 12.9 mmol, 3.0 equiv) およびDBU (1934 μＬ, 12.9 mmol, 3.0 equiv) を加え室温で1時間30分攪拌した。反応液にCH₃CN (200 mL) を加えて析出した沈殿物をろ過し、CH₃CN懸洗を2回行い化合物5 (4.77 g, 98.4%) を得た。
化合物6の合成

Compound 4 (5.65 g, 4.31 mmol) was dissolved in THF (40 mL), and PhSH (1328 μL, 12.9 mmol, 3.0 equiv) and DBU (1934 μL, 12.9 mmol, 3.0 equiv) were added for 1 hour 30 minutes at room temperature. Stir. CH ₃ CN (200 mL) was added to the reaction solution, and the deposited precipitate was filtered, and CH ₃ CN was washed twice to obtain Compound 5 (4.77 g, 98.4%).
Synthesis of compound 6

化合物5 (7.49 g, 6.65 mmol) をTHF (70 mL) に溶解し、Fmoc-Leu-OH (3.53 g, 9.98 mmol, 1.5 equiv)、HOAt (2.27 g, 16.7 mmol, 2.5 equiv)、HATU (6.32 g, 16.6 mmol, 2.5 equiv) およびDIPEA (5800 μＬ, 33.3 mmol, 5.0 equiv) を加えて室温で6分間攪拌した。DIPEA (580 μＬ, 3.33 mmol, 0.5 equiv) を加えて8分間攪拌した。さらに、DIPEA (580 μＬ, 3.33 mmol, 0.5 equiv) を加えて1時間攪拌した。沈殿物をろ過し、ろ液を減圧下留去した。化合物4の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (n-hexane/EtOAc = 100 : 0 - 85 : 15) に付し、化合物6 (9.10 g, 93.7%) を得た。
化合物7の合成

Compound 5 (7.49 g, 6.65 mmol) was dissolved in THF (70 mL) and Fmoc-Leu-OH (3.53 g, 9.98 mmol, 1.5 equiv), HOAt (2.27 g, 16.7 mmol, 2.5 equiv), HATU (6.32 g, 16.6 mmol, 2.5 equiv) and DIPEA (5800 μL, 33.3 mmol, 5.0 equiv) were added, and the mixture was stirred at room temperature for 6 minutes. DIPEA (580 μL, 3.33 mmol, 0.5 equiv) was added and stirred for 8 minutes. Furthermore, DIPEA (580 μL, 3.33 mmol, 0.5 equiv) was added and stirred for 1 hour. The precipitate was filtered and the filtrate was distilled off under reduced pressure. The residue obtained by post-treatment equivalent to the synthesis of compound 4 was subjected to silica gel column chromatography (n-hexane / EtOAc = 100: 0-85: 15) to obtain compound 6 (9.10 g, 93.7%). .
Synthesis of compound 7

化合物6 (1.05g, 0.721 mmol) をCH₂Cl₂ (36 mL) に溶解し、TFE (3.6 mL) およびTFA (360 μＬ) を加えて室温で40分間攪拌した。沈殿物をろ過し、ろ液を減圧下留去した。残渣にCH₂Cl₂を加えて水で3回洗浄し、有機層を飽和食塩水で洗浄、無水MgSO₄で乾燥、ろ過し、ろ液を減圧下留去した。残渣をシリカゲルカラムクロマトグラフィー (CH₂Cl₂/MeOH, 100 : 0 - 90 :10) に付し、化合物7 (478 mg, 92.2%) を得た。
化合物8の合成

Compound 6 (1.05 g, 0.721 mmol) was dissolved in CH ₂ Cl ₂ (36 mL), TFE (3.6 mL) and TFA (360 μL) were added, and the mixture was stirred at room temperature for 40 minutes. The precipitate was filtered and the filtrate was distilled off under reduced pressure. CH ₂ Cl ₂ was added to the residue and washed with water three times. The organic layer was washed with saturated brine, dried over anhydrous MgSO ₄ and filtered, and the filtrate was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography (CH ₂ Cl ₂ / MeOH, 100: 0 to 90:10) to obtain Compound 7 (478 mg, 92.2%).
Synthesis of compound 8

化合物7 (4.08 g, 5.66 mmol, 1.2 equiv) をTHF (70 mL) に溶解し、化合物29（合成方法は後で記載する） (5.52 g, 4.72 mmol)、DIPCI (1104 μＬ, 7.09 mmol, 1.5 equiv)、HOAt (963 mg, 7.08 mmol, 1.5 equiv)、およびDIPEA (4940 μＬ, 28.4 mmol, 6.0 equiv) を加えて室温で3時間攪拌した。HOAt (325 mg, 2.39 mmol, 0.5 equiv) およびDIPCI (368 μＬ, 2.36 mmol, 0.5 equiv) を加えて2時間攪拌した。溶媒を減圧下留去した後、化合物２の合成と同等の後処理を行い化合物8 (8.54 g, 98.5%) を得た。
化合物9の合成

Compound 7 (4.08 g, 5.66 mmol, 1.2 equiv) was dissolved in THF (70 mL), and compound 29 (synthesis method will be described later) (5.52 g, 4.72 mmol), DIPCI (1104 μL, 7.09 mmol, 1.5 equiv), HOAt (963 mg, 7.08 mmol, 1.5 equiv), and DIPEA (4940 μL, 28.4 mmol, 6.0 equiv) were added and stirred at room temperature for 3 hours. HOAt (325 mg, 2.39 mmol, 0.5 equiv) and DIPCI (368 μL, 2.36 mmol, 0.5 equiv) were added and stirred for 2 hours. After the solvent was distilled off under reduced pressure, the post-treatment equivalent to the synthesis of Compound 2 was performed to obtain Compound 8 (8.54 g, 98.5%).
Synthesis of compound 9

化合物8 (8.54 g, 4.65 mmol) をTHF (93 mL) に溶解し、piperdine (891 μＬ, 8.37 mmol, 1.8 equiv) およびDBU (904 μＬ, 6.04 mmol, 1.3 equiv) を加えて室温で5分間攪拌した。化合物2の合成と同等の後処理を行い脱Fmoc体 (8.31 g) を得た。

Compound 8 (8.54 g, 4.65 mmol) is dissolved in THF (93 mL), piperdine (891 μL, 8.37 mmol, 1.8 equiv) and DBU (904 μL, 6.04 mmol, 1.3 equiv) are added and stirred at room temperature for 5 minutes. did. The post-process equivalent to the synthesis | combination of the compound 2 was performed, and the de-Fmoc body (8.31 g) was obtained.

脱Fmoc体 (8.31 g) をTHF (93 mL) に溶解し、Fmoc-Tyr(tBu)-OH (3.21 g, 6.99 mmol, 1.5 equiv)、HOAt (1.59 g, 11.7 mmol, 2.5 equiv)、HATU (4.42 g, 11.6 mmol, 2.5 equiv)、およびDIPEA (4860 μＬ, 27.9 mmol, 6.0 equiv) を加えて室温で40分間攪拌した。化合物４の合成と同等の後処理を行い化合物9 (9.05 g, 94.7%) を得た。
化合物10の合成

De-Fmoc form (8.31 g) was dissolved in THF (93 mL) and Fmoc-Tyr (tBu) -OH (3.21 g, 6.99 mmol, 1.5 equiv), HOAt (1.59 g, 11.7 mmol, 2.5 equiv), HATU ( 4.42 g, 11.6 mmol, 2.5 equiv) and DIPEA (4860 μL, 27.9 mmol, 6.0 equiv) were added, and the mixture was stirred at room temperature for 40 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 9 (9.05 g, 94.7%) was obtained.
Synthesis of compound 10

化合物9 (4.17 g, 2.03 mmol) をTHF (41 mL) に溶解し、piperidine (389 μＬ, 3.65 mmol, 1.8 equiv) およびDBU (395 μＬ, 2.63 mmol, 1.3 equiv) 加えて室温で5分間攪拌した。化合物2の合成と同等の後処理を行い脱Fmoc体 (3.92 g) を得た。

Compound 9 (4.17 g, 2.03 mmol) was dissolved in THF (41 mL), piperidine (389 μL, 3.65 mmol, 1.8 equiv) and DBU (395 μL, 2.63 mmol, 1.3 equiv) were added, and the mixture was stirred at room temperature for 5 minutes. . The post-process equivalent to the synthesis | combination of the compound 2 was performed, and the de-Fmoc body (3.92 g) was obtained.

脱Fmoc体 (3.92 g) をTHF (41 mL) に溶解し、Fmoc-Gln(Trt)-OH (1.86 g, 3.04 mmol, 1.5 equiv)、HATU (1.93 g, 5.07 mmol, 2.5 equiv)、HOAt (691 mg, 5.07 mmol, 2.5 equiv)、およびDIPEA (2120 μＬ, 12.2 mmol, 6.0 equiv) を加えて室温で30分間攪拌した。化合物4の合成と同等の後処理を行い化合物10 (4.83 g, 98.0%) を得た。
化合物11の合成

De-Fmoc form (3.92 g) was dissolved in THF (41 mL), and Fmoc-Gln (Trt) -OH (1.86 g, 3.04 mmol, 1.5 equiv), HATU (1.93 g, 5.07 mmol, 2.5 equiv), HOAt ( 691 mg, 5.07 mmol, 2.5 equiv) and DIPEA (2120 μL, 12.2 mmol, 6.0 equiv) were added and stirred at room temperature for 30 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 10 (4.83 g, 98.0%) was obtained.
Synthesis of compound 11

化合物10 (4.83 g, 1.99 mmol) をTHF (40 mL) に溶解し、piperidine (382 μＬ, 3.59 mmol, 1.8 equiv) およびDBU (388 μＬ, 2.59 mmol, 1.3 equiv) 加えて室温で5分間攪拌した。化合物2の合成と同等の後処理を行い脱Fmoc体 (4.63 g) を得た。

Compound 10 (4.83 g, 1.99 mmol) was dissolved in THF (40 mL), piperidine (382 μL, 3.59 mmol, 1.8 equiv) and DBU (388 μL, 2.59 mmol, 1.3 equiv) were added, and the mixture was stirred at room temperature for 5 minutes. . The post-process equivalent to the synthesis | combination of the compound 2 was performed, and the de-Fmoc body (4.63 g) was obtained.

脱Fmoc体 (4.63 g) をTHF (40 mL) に溶解し、Fmoc-Ser(tBu)-OH (1.15 g, 2.99 mmol, 1.5 equiv)、HATU (1.89 g, 4.98 mmol, 2.5 equiv)、HOAt (676 mg, 4.98 mmol, 2.5 equiv)、およびDIPEA (2082 μＬ, 11.9 mmol, 6.0 equiv) を加えて室温で45分間攪拌した。化合物4の合成と同等の後処理を行い化合物11 (5.49 g) を得た。
化合物12の合成

De-Fmoc form (4.63 g) was dissolved in THF (40 mL) and Fmoc-Ser (tBu) -OH (1.15 g, 2.99 mmol, 1.5 equiv), HATU (1.89 g, 4.98 mmol, 2.5 equiv), HOAt ( 676 mg, 4.98 mmol, 2.5 equiv) and DIPEA (2082 μL, 11.9 mmol, 6.0 equiv) were added and stirred at room temperature for 45 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 11 (5.49 g) was obtained.
Synthesis of Compound 12

化合物11 (5.49 g) をTHF (40 mL) に溶解し、piperidine (382 μＬ, 3.59 mmol, 1.8 equiv) およびDBU (388 μＬ, 2.59 mmol, 1.3 equiv) 加えて室温で5分間攪拌した。化合物2の合成と同等の後処理を行い脱Fmoc体 (4.65 g, 98.0% from 10) を得た。

Compound 11 (5.49 g) was dissolved in THF (40 mL), piperidine (382 μL, 3.59 mmol, 1.8 equiv) and DBU (388 μL, 2.59 mmol, 1.3 equiv) were added, and the mixture was stirred at room temperature for 5 minutes. The post-process equivalent to the synthesis | combination of the compound 2 was performed, and the de-Fmoc body (4.65 g, 98.0% from 10) was obtained.

脱Fmoc体 (4.65 g, 1.95 mmol) をTHF (39 mL) に溶解し、Fmoc-Trp(Boc)-OH (1.54 g, 2.92 mmol, 1.5 equiv)、HOAt (398 mg, 2.92 mmol, 1.5 equiv)、HATU (1.11 g, 2.92 mmol, 1.5 equiv)、およびDIPEA (1698 μＬ, 9.75 mmol, 5.0 equiv) を加えて室温で70分間攪拌した。化合物4の合成と同等の後処理を行い化合物12 (5.32 g, 95.4%) を得た。
化合物13の合成

De-Fmoc form (4.65 g, 1.95 mmol) is dissolved in THF (39 mL), Fmoc-Trp (Boc) -OH (1.54 g, 2.92 mmol, 1.5 equiv), HOAt (398 mg, 2.92 mmol, 1.5 equiv) , HATU (1.11 g, 2.92 mmol, 1.5 equiv) and DIPEA (1698 μL, 9.75 mmol, 5.0 equiv) were added and stirred at room temperature for 70 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 12 (5.32 g, 95.4%) was obtained.
Synthesis of compound 13

化合物12 (5.32 g, 1.86 mmol) をTHF (37.2 mL) に溶解し、piperidine (372 μＬ, 3.50 mmol, 1.9 equiv) およびDBU (372 μＬ, 2.49 mmol, 1.3 equiv) 加えて室温で5分間攪拌した。化合物2の合成と同等の後処理を行い脱Fmoc体 (4.96 g) を得た。

Compound 12 (5.32 g, 1.86 mmol) was dissolved in THF (37.2 mL), piperidine (372 μL, 3.50 mmol, 1.9 equiv) and DBU (372 μL, 2.49 mmol, 1.3 equiv) were added, and the mixture was stirred at room temperature for 5 minutes. . The post-process equivalent to the synthesis | combination of the compound 2 was performed, and the de-Fmoc body (4.96 g) was obtained.

脱Fmoc体 (4.96 g) をTHF (12.6 mL) に溶解し、TBAF (1.0 M solution in THF, 6.0 mL, 6.00 mmol, 3.2 equiv) を加えて室温で19時間攪拌した。減圧下溶媒を留去した後、残渣にCH₂Cl₂を加えて1 N HCl、水、飽和食塩水で洗浄し、有機層を無水MgSO₄で乾燥しろ過後、ろ液を減圧下留去した。残渣をシリカゲルカラムクロマトグラフィー (CH₂Cl₂/THF, 100 : 0 - 75 : 25) に付し、化合物13 (2.02 g, 45.3%) を得た。
化合物29の合成

The de-Fmoc form (4.96 g) was dissolved in THF (12.6 mL), TBAF (1.0 M solution in THF, 6.0 mL, 6.00 mmol, 3.2 equiv) was added, and the mixture was stirred at room temperature for 19 hours. After evaporating the solvent under reduced pressure, CH ₂ Cl ₂ was added to the residue, and the residue was washed with 1 N HCl, water, and saturated brine. The organic layer was dried over anhydrous MgSO ₄ and filtered, and the filtrate was evaporated under reduced pressure. . The residue was subjected to silica gel column chromatography (CH ₂ Cl ₂ / THF, 100: 0 to 75:25) to give compound 13 (2.02 g, 45.3%).
Synthesis of compound 29

3,4,5-トリオクタデシルオキシベンジルアルコール （「ＫａＯＨ」と表記する）(20.1 g, 22.0 mmol) をCH₂Cl₂ (220 mL) に溶解し、Fmoc-Tyr(tBu)-OH (15.2 g, 33.0 mmol, 1.5 equiv)、DIPCI (6856 μＬ, 44.0 mmol, 2.0 equiv)、およびDMAP (26.9 mg, 0.220 mmol, 0.01 equiv) を加えて室温で35分間攪拌した。化合物1の合成と同等の後処理を行い化合物28 (29.9 g, quant. y.) を得た。

3,4,5-Trioctadecyloxybenzyl alcohol (referred to as “KaOH”) (20.1 g, 22.0 mmol) was dissolved in CH ₂ Cl ₂ (220 mL) and Fmoc-Tyr (tBu) -OH (15.2 g , 33.0 mmol, 1.5 equiv), DIPCI (6856 μL, 44.0 mmol, 2.0 equiv), and DMAP (26.9 mg, 0.220 mmol, 0.01 equiv) were added and stirred at room temperature for 35 minutes. The post-process equivalent to the synthesis | combination of the compound 1 was performed, and the compound 28 (29.9 g, quant. Y.) Was obtained.

化合物28 (9.58 g, 7.07 mmol) をTHF (141 mL) に溶解し、piperidine (1260 μＬ, 12.7 mmol, 1.8 equiv) およびDBU (1374 μＬ, 9.19 mmol, 1.3 equiv) を加えて室温で5分間攪拌した。TLCにて原料消失を確認後、化合物2の合成と同等の後処理を行い化合物29 (8.28 g, quant. y.) を得た。
１−２．原料である化合物３８の合成
化合物30の合成

Compound 28 (9.58 g, 7.07 mmol) is dissolved in THF (141 mL), piperidine (1260 μL, 12.7 mmol, 1.8 equiv) and DBU (1374 μL, 9.19 mmol, 1.3 equiv) are added and stirred at room temperature for 5 minutes. did. After confirming disappearance of the raw materials by TLC, post-treatment equivalent to the synthesis of compound 2 was performed to obtain compound 29 (8.28 g, quant. Y.).
1-2. Synthesis of compound 38 as raw material Synthesis of compound 30

3,4,5-トリオクタデシルオキシベンジルアルコール (3.00 g, 3.28 mmol) をCH₂Cl₂ (32.8 mL) に溶解し、Fmoc-Lys(Mtt)-OH (3.08 g, 4.93 mmol, 1.5 equiv)、DIPCI (1023 μＬ, 6.57 mmol, 2.0 equiv)、およびDMAP (4.0 mg, 0.0328 mmol, 0.01 equiv) を加えて室温で42分間攪拌した。化合物1の合成と同等の後処理を行い化合物30 (5.32 g) を得た。
化合物31の合成

3,4,5-trioctadecyloxybenzyl alcohol (3.00 g, 3.28 mmol) was dissolved in CH ₂ Cl ₂ (32.8 mL) and Fmoc-Lys (Mtt) -OH (3.08 g, 4.93 mmol, 1.5 equiv), DIPCI (1023 μL, 6.57 mmol, 2.0 equiv) and DMAP (4.0 mg, 0.0328 mmol, 0.01 equiv) were added and stirred at room temperature for 42 minutes. The post-process equivalent to the synthesis | combination of the compound 1 was performed, and the compound 30 (5.32 g) was obtained.
Synthesis of compound 31

化合物30 (5.32 g) をCH₂Cl₂ (32.8 mL) に溶解し、TIS (3284 μＬ) およびTFA (985.2 μＬ) を加えて室温で12分間攪拌した。TIS (3284 μＬ) を追加し更に53分間攪拌した。TFA (985.2 μＬ) を追加して更に16分間攪拌した。化合物4の合成と同等の後処理を行い化合物31 (4.48 g) を得た。
化合物32の合成

Compound 30 (5.32 g) was dissolved in CH ₂ Cl ₂ (32.8 mL), TIS (3284 μL) and TFA (985.2 μL) were added, and the mixture was stirred at room temperature for 12 minutes. TIS (3284 μL) was added, and the mixture was further stirred for 53 minutes. TFA (985.2 μL) was added, and the mixture was further stirred for 16 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 31 (4.48 g) was obtained.
Synthesis of compound 32

化合物31 (4.48 g) をCH₂Cl₂ (65.7 mL) に溶解し、DIPEA (1258 μＬ, 7.22 mmol, 2.2 equiv) およびNsCl (873 mg, 3.94 mmol, 1.2 equiv) を加えて室温で37分間攪拌した。化合物3の合成と同等の後処理を行い化合物32 (4.75 g, quant. y.) を得た。
化合物33の合成

Compound 31 (4.48 g) was dissolved in CH ₂ Cl ₂ (65.7 mL), DIPEA (1258 μL, 7.22 mmol, 2.2 equiv) and NsCl (873 mg, 3.94 mmol, 1.2 equiv) were added and stirred at room temperature for 37 minutes. did. The post-process equivalent to the synthesis | combination of the compound 3 was performed, and the compound 32 (4.75 g, quant. Y.) Was obtained.
Synthesis of compound 33

化合物32 (4.38 g, 3.02 mmol) をCH₂Cl₂ (27 mL) に溶解し、TIS (6.0 mL) およびTFA (27 mL) を加えて室温で 2時間攪拌した。減圧下溶媒を留去し、残渣にCH₂Cl₂加えて希釈し水で3回、飽和食塩水で洗浄し、有機層を無水MgSO4で乾燥、ろ過し、ろ液を減圧下留去した。残渣をシリカゲルカラムクロマトグラフィー (CH₂Cl₂/MeOH, 100 : 0 - 85 : 15) に付し、化合物33 (1.42 g, 84.8%) を得た。
化合物34の合成

Compound 32 (4.38 g, 3.02 mmol) was dissolved in CH ₂ Cl ₂ (27 mL), TIS (6.0 mL) and TFA (27 mL) were added, and the mixture was stirred at room temperature for 2 hr. The solvent was distilled off under reduced pressure, CH ₂ Cl _{2 was} added to the residue, diluted, washed three times with water and with saturated brine, the organic layer was dried over anhydrous MgSO 4 and filtered, and the filtrate was evaporated under reduced pressure. The residue was subjected to silica gel column chromatography (CH ₂ Cl ₂ / MeOH, 100: 0 to 85:15) to obtain Compound 33 (1.42 g, 84.8%).
Synthesis of Compound 34

2,4-ジドコソキシベンジルアルコール (2.09 g, 2.76 mmol) をCH₂Cl₂ (27.6 mL) に溶解し、化合物33 (2.29 g, 4.14 mmol, 1.5 equiv)、DIPCI (859 μＬ, 5.51 mmol, 2.0 equiv)、およびDMAP (3.4 mg, 0.0278 mmol, 0.01 equiv) を加えて室温で45分間攪拌した。化合物1と同等の後処理を行い化合物34 (4.13 g, quant. y.) を得た。
化合物35の合成

2,4-Didocosoxybenzyl alcohol (2.09 g, 2.76 mmol) was dissolved in CH ₂ Cl ₂ (27.6 mL), compound 33 (2.29 g, 4.14 mmol, 1.5 equiv), DIPCI (859 μL, 5.51 mmol, 2.0 equiv) and DMAP (3.4 mg, 0.0278 mmol, 0.01 equiv) were added and stirred at room temperature for 45 minutes. The post-process equivalent to the compound 1 was performed, and the compound 34 (4.13 g, quant. Y.) Was obtained.
Synthesis of compound 35

化合物34 (4.13) をTHF (55.1 mL) に溶解し、piperidine (491 μＬ) およびDBU (536 μＬ) を加えて室温で5分間攪拌した。TLCにて原料消失を確認後、化合物2の合成と同等の後処理を行い化合物35 (3.00 g, 98.2%) を得た。
化合物36の合成

Compound 34 (4.13) was dissolved in THF (55.1 mL), piperidine (491 μL) and DBU (536 μL) were added, and the mixture was stirred at room temperature for 5 minutes. After confirming disappearance of the raw material by TLC, the post-treatment equivalent to the synthesis of Compound 2 was performed to obtain Compound 35 (3.00 g, 98.2%).
Synthesis of compound 36

化合物35 (332 mg, 0.300 mmol) をTHF (6 mL) に溶解し、Fmoc-Tyr(tBu)-OH (207 mg, 0.450 mmol, 1.5 equiv)、HOAt (61.3 mg, 0.450 mmol, 1.5 equiv)、HATU (171 mg, 0.450 mmol, 1.5 equiv) およびDIPEA (261 μＬ, 1.50 mmol, 5.0 equiv) を加えて室温で53分間攪拌した。化合物4の合成と同等の後処理を行い化合物36 (441 mg, 97.3%) を得た。化合物37の合成

Compound 35 (332 mg, 0.300 mmol) was dissolved in THF (6 mL), Fmoc-Tyr (tBu) -OH (207 mg, 0.450 mmol, 1.5 equiv), HOAt (61.3 mg, 0.450 mmol, 1.5 equiv), HATU (171 mg, 0.450 mmol, 1.5 equiv) and DIPEA (261 μL, 1.50 mmol, 5.0 equiv) were added and stirred at room temperature for 53 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 36 (441 mg, 97.3%) was obtained. Synthesis of compound 37

化合物36 (2.73 g, 1.81 mmol) をTHF (36.1 mL) に溶解し、piperidine (322 μＬ) およびDBU (351 μＬ) を加えて室温で5分間攪拌した。TLCにて原料消失を確認後、化合物2の合成と同等の後処理を行い化合物37 (2.25 g, 94.1%) を得た。
化合物38の合成

Compound 36 (2.73 g, 1.81 mmol) was dissolved in THF (36.1 mL), piperidine (322 μL) and DBU (351 μL) were added, and the mixture was stirred at room temperature for 5 minutes. After confirming disappearance of the raw material by TLC, the post-treatment equivalent to the synthesis of Compound 2 was performed to obtain Compound 37 (2.25 g, 94.1%).
Synthesis of compound 38

化合物37 (266 mg, 0.201 mmol) をTHF (4 mL) に溶解し、ミリスチン酸 (68.9 mg, 0.302 mmol, 1.5 equiv)、HOAt (41.4 mg, 0.304 mmol, 1.5 equiv)、HATU (114 mg, 0.301 mmol, 1.5 equiv)、およびDIPEA (174 μＬ, 0.999 mmol, 5.0 equiv) を加えて室温で1時間50分間攪拌した。化合物4の合成と同等の後処理を行い化合物38 (289 mg, 95.5%) を得た。
１−３．中間体である化合物１４の合成

Compound 37 (266 mg, 0.201 mmol) was dissolved in THF (4 mL), myristic acid (68.9 mg, 0.302 mmol, 1.5 equiv), HOAt (41.4 mg, 0.304 mmol, 1.5 equiv), HATU (114 mg, 0.301 mmol, 1.5 equiv) and DIPEA (174 μL, 0.999 mmol, 5.0 equiv) were added, and the mixture was stirred at room temperature for 1 hour and 50 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 38 (289 mg, 95.5%) was obtained.
1-3. Synthesis of intermediate compound 14

化合物13 (265 mg, 0.111 mmol)、化合物38 (289 mg, 0.192 mmol, 1.7 equiv)、およびPPh₃ (118 mg, 0.450 mmol, 4.1 equiv) をTHF (11 mL) に溶解し、DEAD (201 μＬ, 0.443 mmol, 4.0 equiv) を加えて室温で61分間攪拌した。PPh₃ (121 mg, 0.461 mmol, 4.2 equiv) およびDEAD (201 μＬ, 0.0.443 mmol, 4.0 equiv) を加えて59分間攪拌した。化合物３の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (CH₂CH₂/THF, 100 : 0 - 88 : 12) に付し、光延反応物 (132 mg, 30.6%) を得た。

Compound 13 (265 mg, 0.111 mmol), Compound 38 (289 mg, 0.192 mmol, 1.7 equiv), and PPh ₃ (118 mg, 0.450 mmol, 4.1 equiv) were dissolved in THF (11 mL) and DEAD (201 μL , 0.443 mmol, 4.0 equiv) was added and stirred at room temperature for 61 minutes. PPh ₃ (121 mg, 0.461 mmol, 4.2 equiv) and DEAD (201 μL, 0.0.443 mmol, 4.0 equiv) were added and stirred for 59 minutes. The residue obtained by post-treatment equivalent to the synthesis of compound 3 was subjected to silica gel column chromatography (CH ₂ CH ₂ / THF, 100: 0-88: 12) to give Mitsunobu reaction product (132 mg, 30.6%). Obtained.

光延反応物 (132 mg, 0.0340 mmol) をCH₂Cl₂ (3.4 mL) に溶解し、TFE (340 μＬ) およびTFA (34.0 μＬ) を加えて室温で70分間攪拌した。沈殿物をセライトろ過し、減圧下溶媒を留去した。化合物4の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (CHCl₃/MeOH, 100 : 0 - 95 : 5) に付し、本発明の中間体である化合物14 (75.1 mg, 70.3%) を得た
（実施例２）ＷＰ９ＱＹペプチド模倣体（Bdev-7）の合成
化合物15の合成

Mitsunobu reaction product (132 mg, 0.0340 mmol) was dissolved in CH ₂ Cl ₂ (3.4 mL), TFE (340 μL) and TFA (34.0 μL) were added, and the mixture was stirred at room temperature for 70 minutes. The precipitate was filtered through celite, and the solvent was distilled off under reduced pressure. The residue obtained by post-treatment equivalent to the synthesis of Compound 4 was subjected to silica gel column chromatography (CHCl ₃ / MeOH, 100: 0 to 95: 5) to give Compound 14 (75.1 mg, intermediate) of the present invention. Example 2) Synthesis of WP9QY Peptidomimetic (Bdev-7) Synthesis of Compound 15

化合物14 (75.1 mg, 0.0239 mmol)、HOAt (4.2 mg, 0.0309 mmol, 1.3 equiv)、およびHATU (11.2 mg, 0.0295 mmol, 1.2 equiv) をTHF (4780 μＬ) に溶解し、DIPEA (20.8 μＬ, 0.119 mmol, 5.0 equiv) を加えて室温で17時間40分間攪拌した。化合物4の合成と同等の後処理を行い化合物15 (64.5 mg, 86.6%) を得た。
化合物16の合成

Compound 14 (75.1 mg, 0.0239 mmol), HOAt (4.2 mg, 0.0309 mmol, 1.3 equiv), and HATU (11.2 mg, 0.0295 mmol, 1.2 equiv) were dissolved in THF (4780 μL) and DIPEA (20.8 μL, 0.119 mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 17 hours and 40 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 15 (64.5 mg, 86.6%) was obtained.
Synthesis of Compound 16

化合物15 (64.5 mg, 0.0207 mmol) をTHF (414 μＬ) に溶解し、PhSH (6.38 μＬ, 0.0621 mmol, 3.0 equiv) およびDBU (9.29 μＬ, 0.0621 mmol, 3.0 equiv) を加えて室温で1時間38分間攪拌した。PhSH (6.38 μＬ, 0.0621 mmol, 3.0 equiv) およびDBU (9.29 μＬ, 0.0621 mmol, 3.0 equiv) を加えて室温で3時間29分間攪拌した。反応液に濃塩酸 (10 μＬ) を加えて減圧下溶媒を留去した。化合物4の合成と同等の後処理を行い化合物16 (55.6 mg, 91.3%) を得た。
Bdev-7の合成

Compound 15 (64.5 mg, 0.0207 mmol) was dissolved in THF (414 μL), and PhSH (6.38 μL, 0.0621 mmol, 3.0 equiv) and DBU (9.29 μL, 0.0621 mmol, 3.0 equiv) were added for 1 hour at room temperature 38 Stir for minutes. PhSH (6.38 μL, 0.0621 mmol, 3.0 equiv) and DBU (9.29 μL, 0.0621 mmol, 3.0 equiv) were added, and the mixture was stirred at room temperature for 3 hours and 29 minutes. Concentrated hydrochloric acid (10 μL) was added to the reaction solution, and the solvent was distilled off under reduced pressure. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 16 (55.6 mg, 91.3%) was obtained.
Synthesis of Bdev-7

化合物16 (21.8 mg, 0.0743 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (743 μＬ) を加えて室温で3時間攪拌した。沈殿物をろ過し、ろ液を減圧下留去した。残渣にIPEを加え沈殿物を析出させ、遠心分離後の残渣をHPLC精製し本発明の架橋ペプチドBdev-7 (2.8 mg, 2.6%) を得た。HRMS m/z [M + H]⁺: calcd for C₇₈H₁₁₁N₁₂O₁₆: 1471.8241, found 1472.1734。
下記のペプチド配列構造を有する他のＷＰ９ＱＹペプチド模倣体を合成した。

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (743 μL) was added to compound 16 (21.8 mg, 0.0743 mmol), and the mixture was stirred at room temperature for 3 hours. The precipitate was filtered and the filtrate was distilled off under reduced pressure. IPE was added to the residue to precipitate a precipitate, and the residue after centrifugation was purified by HPLC to obtain the cross-linked peptide Bdev-7 (2.8 mg, 2.6%) of the present invention. HRMS m / z [M + H] ⁺ : calcd for C ₇₈ H ₁₁₁ N ₁₂ O ₁₆ : 1471.8241, found 1472.1734.
Another WP9QY peptidomimetic having the following peptide sequence structure was synthesized.

合成したそれぞれの化合物のＺ₁〜Ｚ₃は、以下の表２に記載の通りである。

Z _{1 to} Z ₃ of each synthesized compound are as shown in Table 2 below.

Example 3 Synthesis of Intermediate (Compound 19) for Synthesis of Bdev-3, -5, -6, -12 and -13
A schematic diagram of the synthesis of intermediates for the synthesis of Bdev-3, -5, -6, -12 and -13 is shown below.

それぞれの合成は、以下のようにして行った。
化合物39の合成

Each synthesis was performed as follows.
Synthesis of compound 39

化合物37 (746 mg, 0.562 mmol) をCH₂Cl₂ (11 mL) に溶解し、Et₃N (160 μＬ, 1.14 mmol, 2.0 equiv) およびAc₂O (100 μＬ, 1.06 mmol, 1.9 equiv) を加えて室温で52分間攪拌した。化合物3の合成と同等の後処理を行い化合物39 (698 mg, 0.524 mmol, 93.2%) を得た。
化合物40の合成

Compound 37 (746 mg, 0.562 mmol) was dissolved in CH ₂ Cl ₂ (11 mL), and Et ₃ N (160 μL, 1.14 mmol, 2.0 equiv) and Ac ₂ O (100 μL, 1.06 mmol, 1.9 equiv) were dissolved. In addition, the mixture was stirred at room temperature for 52 minutes. The post-process equivalent to the synthesis | combination of the compound 3 was performed, and the compound 39 (698 mg, 0.524 mmol, 93.2%) was obtained.
Synthesis of compound 40

化合物35 (775 mg, 0.700 mmol) をTHF (14 mL) に溶解し、Boc-Tyr(tBu)-OH (354 mg, 1.05 mmol, 1.5 equiv)、HOAt (143 mg, 1.05 mmol, 1.5 equiv)、HATU (399 mg, 1.05 mmol, 1.5 equiv) およびDIPEA (610 μＬ, 3.50 mmol, 5.0 equiv) を加えて室温で42分間攪拌した。化合物4の合成と同等の後処理を行い化合物40 (935 mg, 96.1%) を得た。
化合物17の合成

Compound 35 (775 mg, 0.700 mmol) was dissolved in THF (14 mL), Boc-Tyr (tBu) -OH (354 mg, 1.05 mmol, 1.5 equiv), HOAt (143 mg, 1.05 mmol, 1.5 equiv), HATU (399 mg, 1.05 mmol, 1.5 equiv) and DIPEA (610 μL, 3.50 mmol, 5.0 equiv) were added and stirred at room temperature for 42 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 40 (935 mg, 96.1%) was obtained.
Synthesis of compound 17

化合物13 (944 mg, 0.394 mmol)、化合物39 (635 mg, 0.477 mmol, 1.2 equiv)、およびPPh₃ (211 mg, 0.804 mmol, 2.0 equiv) をTHF (40 mL) に溶解し、DEAD (357 μＬ, 0.787 mmol, 2.0 equiv) を加えて室温で3時間7分攪拌した。次に、PPh₃ (208 mg, 0.793 mmol, 2.0 equiv) を加えて33分間攪拌した。さらに、DEAD (357 μＬ, 0.787 mmol, 2.0 equiv) を加えて1時間50分攪拌した。化合物4の合成と同等の後処理を行い得た残渣 (1.573 g) を次反応に用いた。

Compound 13 (944 mg, 0.394 mmol), Compound 39 (635 mg, 0.477 mmol, 1.2 equiv), and PPh ₃ (211 mg, 0.804 mmol, 2.0 equiv) were dissolved in THF (40 mL) and DEAD (357 μL , 0.787 mmol, 2.0 equiv) was added, and the mixture was stirred at room temperature for 3 hours and 7 minutes. Next, PPh ₃ (208 mg, 0.793 mmol, 2.0 equiv) was added and stirred for 33 minutes. Furthermore, DEAD (357 μL, 0.787 mmol, 2.0 equiv) was added and stirred for 1 hour and 50 minutes. The residue (1.573 g) obtained by performing post-treatment equivalent to the synthesis of compound 4 was used in the next reaction.

残渣 (1.573 g) をCH₂Cl₂ (47.7 mL) に溶解し、TFE (4770 μＬ) およびTFA (477 μＬ) を加えて室温で1時間攪拌した。沈殿物をセライトろ過し、ろ液にDIPEA (1050 μＬ) を加えた後、減圧下溶媒を留去した。化合物4の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (CHCl₃/MeOH, 100 : 0 - 90 : 10 およびCHCl₃/EtOH, 100 : 0 - 93 : 7) に付し、本発明の中間体である化合物17 (651 mg, 55.7%) を得た。
化合物18の合成

The residue (1.573 g) was dissolved in CH ₂ Cl ₂ (47.7 mL), TFE (4770 μL) and TFA (477 μL) were added, and the mixture was stirred at room temperature for 1 hour. The precipitate was filtered through Celite, DIPEA (1050 μL) was added to the filtrate, and the solvent was evaporated under reduced pressure. The residue obtained by post-treatment equivalent to the synthesis of Compound 4 was subjected to silica gel column chromatography (CHCl ₃ / MeOH, 100: 0-90:10 and CHCl ₃ / EtOH, 100: 0-93: 7), Compound 17 (651 mg, 55.7%) which was an intermediate of the present invention was obtained.
Synthesis of compound 18

化合物17 (485 mg, 0.163 mmol)、HOAt (26.8 mg, 0.197 mmol, 1.2 equiv)、およびHATU (74.8 mg, 0.197 mmol, 1.2 equiv) をTHF (32.6 mL) に溶解し、DIPEA (142 μＬ, 0.815 mmol, 5.0 equiv) を加えて室温で3時間30分間攪拌した。化合物4の合成と同等の後処理を行い化合物18 (462 mg, 96.3%) を得た。
化合物19の合成

Compound 17 (485 mg, 0.163 mmol), HOAt (26.8 mg, 0.197 mmol, 1.2 equiv), and HATU (74.8 mg, 0.197 mmol, 1.2 equiv) were dissolved in THF (32.6 mL) and DIPEA (142 μL, 0.815 mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 3 hours and 30 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 18 (462 mg, 96.3%) was obtained.
Synthesis of compound 19

化合物40 (462 mg, 0.157 mmol) をTHF (1560 μＬ) に溶解し、PhSH (16.0 μＬ, 0.156 mmol, 1.0 equiv) およびDBU (70.0 μＬ, 0.468 mmol, 3.0 equiv) を加えて室温で50分間攪拌した。PhSH (32.0 μＬ, 0.312 mmol, 2.0 equiv) を加えて1時間11分攪拌した。PhSH (48.0 μＬ, 0.467 mmol, 3.0 equiv) およびDBU (70.0 μＬ, 0.468 mmol, 3.0 equiv) を加えて室温で50分間攪拌した。反応液に濃塩酸 (78.0 μＬ) を加えて減圧下溶媒を留去した。化合物４の合成と同等の後処理を行い化合物19 (416 mg, 96.2%) を得た。

Compound 40 (462 mg, 0.157 mmol) is dissolved in THF (1560 μL), PhSH (16.0 μL, 0.156 mmol, 1.0 equiv) and DBU (70.0 μL, 0.468 mmol, 3.0 equiv) are added and stirred at room temperature for 50 minutes. did. PhSH (32.0 μL, 0.312 mmol, 2.0 equiv) was added and stirred for 1 hour and 11 minutes. PhSH (48.0 μL, 0.467 mmol, 3.0 equiv) and DBU (70.0 μL, 0.468 mmol, 3.0 equiv) were added and stirred at room temperature for 50 minutes. Concentrated hydrochloric acid (78.0 μL) was added to the reaction solution, and the solvent was distilled off under reduced pressure. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 19 (416 mg, 96.2%) was obtained.

(Example 4) Synthesis of Bdev-3, -5, -6, -12 and -13 (1) Synthesis of Bdev-6
Bdev-6 was synthesized as follows.

化合物19 (48.2 mg, 0.0174 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (1740 μＬ) を加えて室温で3時間攪拌した。Bdev−７の合成と同等の後処理およびHPLC精製を行いBdev-6 (5.3 mg, 23.4%) を得た。HRMS m/z [M + H]⁺: calcd for C₆₆H₈₇N₁₂O₁₆: 1303.6363, found 1303.9331.
（２）Bdev-12の合成
Bdev-12は、以下の通りにして合成した。
化合物20の合成

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (1740 μL) was added to compound 19 (48.2 mg, 0.0174 mmol), and the mixture was stirred at room temperature for 3 hours. Post-treatment and HPLC purification equivalent to the synthesis of Bdev-7 were performed to obtain Bdev-6 (5.3 mg, 23.4%). HRMS m / z [M + H] ⁺ : calcd for C ₆₆ H ₈₇ N ₁₂ O ₁₆ : 1303.6363, found 1303.9331.
(2) Synthesis of Bdev-12
Bdev-12 was synthesized as follows.
Synthesis of compound 20

化合物19 (149 mg, 0.0540 mmol)、ミリスチン酸 (18.7 mg, 0.0819 mmol, 1.5 equiv)、HOAt (11.1 mg, 0.0816 mmol, 1.5 equiv)、およびHATU (31.0 mg, 0.0815 mmol, 1.5 equiv) をTHF (1080 μＬ) に溶解し、DIPEA (47.0 μＬ, 0.270 mmol, 5.0 equiv) を加えて室温で3時間攪拌した。化合物4の合成と同等の後処理を行い化合物20 (147 mg, 91.5%) を得た。
Bdev-12の合成

Compound 19 (149 mg, 0.0540 mmol), myristic acid (18.7 mg, 0.0819 mmol, 1.5 equiv), HOAt (11.1 mg, 0.0816 mmol, 1.5 equiv), and HATU (31.0 mg, 0.0815 mmol, 1.5 equiv) in THF ( (1080 μL), DIPEA (47.0 μL, 0.270 mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 3 hours. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 20 (147 mg, 91.5%) was obtained.
Synthesis of Bdev-12

化合物20 (147 mg, 0.0494 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (5 mL) を加えて室温で3時間攪拌した。Bdev-7の合成と同等の後処理およびHPLC精製を行いBdev-12 (28.0 mg, 37.4%) を得た。HRMS m/z [M + H]⁺: calcd for C₈₀H₁₁₃N₁₂O₁₇: 1513.8347, found 1514.2531.
（３）Bdev-3の合成
Bdev-3は、以下の通りにして合成した。
化合物21の合成

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (5 mL) was added to compound 20 (147 mg, 0.0494 mmol), and the mixture was stirred at room temperature for 3 hours. Post-treatment and HPLC purification equivalent to the synthesis of Bdev-7 were performed to obtain Bdev-12 (28.0 mg, 37.4%). HRMS m / z [M + H] ⁺ : calcd for C ₈₀ H ₁₁₃ N ₁₂ O ₁₇ : 1513.8347, found 1514.2531.
(3) Synthesis of Bdev-3
Bdev-3 was synthesized as follows.
Synthesis of compound 21

化合物19 (111 mg, 0.0400 mmol)、MeO-PEG-CO₂H（MW：２，０００Ｄａ、Ｉｒｉｓ Ｂｉｏｔｅｃｈ社製） (121 mg, 0.0600 mmol, 1.5 equiv)、HOAt (6.5 mg, 0.0480 mmol, 1.2 equiv)、およびHATU (18.3 mg, 0.0480 mmol, 1.2 equiv) をTHF (2 mL) に溶解し、DIPEA (34.8 μＬ, 0.200 mmol, 5.0 equiv) を加えて室温で53分間攪拌した。DIPEA (34.8 μＬ, 0.200 mmol, 5.0 equiv) を加えて56分間攪拌した。MeO-PEG-CO₂H (121 mg, 0.0600 mmol, 1.5 equiv)、HATU (22.8 mg, 0.0600 mmol, 1.5 equiv)、およびDIPEA (17.4 mL, 0.0999 mmol, 2.5 equiv) を加えて2時間9分攪拌した。減圧下溶媒を留去し、残渣をシリカゲルカラムクロマトグラフィー (CH₂Cl₂/MeOH, 100 : 0 - 90 : 10) に付し、化合物21 (313 mg) を粗生成物として得た。

Compound 19 (111 mg, 0.0400 mmol), MeO-PEG-CO ₂ H (MW: 2,000 Da, manufactured by Iris Biotech) (121 mg, 0.0600 mmol, 1.5 equiv), HOAt (6.5 mg, 0.0480 mmol, 1.2 equiv ) And HATU (18.3 mg, 0.0480 mmol, 1.2 equiv) were dissolved in THF (2 mL), DIPEA (34.8 μL, 0.200 mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 53 minutes. DIPEA (34.8 μL, 0.200 mmol, 5.0 equiv) was added and stirred for 56 minutes. Add MeO-PEG-CO ₂ H (121 mg, 0.0600 mmol, 1.5 equiv), HATU (22.8 mg, 0.0600 mmol, 1.5 equiv), and DIPEA (17.4 mL, 0.0999 mmol, 2.5 equiv) and stir for 2 hours and 9 minutes did. The solvent was distilled off under reduced pressure, and the residue was subjected to silica gel column chromatography (CH ₂ Cl ₂ / MeOH, 100: 0 to 90:10) to obtain compound 21 (313 mg) as a crude product.

Bdev-3 synthesis

化合物21 (147 mg, 0.0494 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (5 mL) を加えて室温で3時間攪拌した。Bdev-7の合成と同等の後処理およびHPLC精製を行いBdev-3 (28.0 mg, 37.4%) を得た。m/z 3140付近を中心に44マス間隔で1価イオン、1640付近を中心に22マス間隔で2価イオンを検出した。
（４）Bdev-5の合成
Bdev-5は、以下の通りにして合成した。
化合物22の合成

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (5 mL) was added to compound 21 (147 mg, 0.0494 mmol), and the mixture was stirred at room temperature for 3 hours. Post-treatment and HPLC purification equivalent to the synthesis of Bdev-7 were performed to obtain Bdev-3 (28.0 mg, 37.4%). Monovalent ions were detected at intervals of 44 masses around m / z 3140, and divalent ions were detected at intervals of 22 masses around 1640.
(4) Synthesis of Bdev-5
Bdev-5 was synthesized as follows.
Synthesis of compound 22

化合物19 (161 mg, 0.0580 mmol) をCH₂Cl₂ (1160 μＬ) に溶解し、Et₃N (15.8 μＬ, 0.112 mmol, 1.9 equiv) およびAc₂O (11.0 μＬ, 0.116 mmol, 2.0 equiv) を加えて室温で40分間攪拌した。化合物4の合成と同等の後処理を行い化合物22 (143 mg, 87.6%) を得た。
Bdev-5の合成

Compound 19 (161 mg, 0.0580 mmol) was dissolved in CH ₂ Cl ₂ (1160 μL), and Et ₃ N (15.8 μL, 0.112 mmol, 1.9 equiv) and Ac ₂ O (11.0 μL, 0.116 mmol, 2.0 equiv) were dissolved. In addition, the mixture was stirred at room temperature for 40 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 22 (143 mg, 87.6%) was obtained.
Synthesis of Bdev-5

化合物22 (143 mg, 0.0508 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (5 mL) を加えて室温で3時間攪拌した。Bdev-7の合成と同等の後処理およびHPLC精製を行いBdev-5 (25.6 mg, 37.4%) を得た。HRMS m/z [M + H]⁺: calcd for C₆₈H₈₈N₁₂O₁₇: 1345.6499, found 1345.5516.

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (5 mL) was added to compound 22 (143 mg, 0.0508 mmol), and the mixture was stirred at room temperature for 3 hours. Post-treatment and HPLC purification equivalent to the synthesis of Bdev-7 were performed to obtain Bdev-5 (25.6 mg, 37.4%). HRMS m / z [M + H] ⁺ : calcd for C ₆₈ H ₈₈ N ₁₂ O ₁₇ : 1345.6499, found 1345.5516.

(5) Synthesis of Bdev-13
Bdev-13 was synthesized as follows.

Bdev-５ (10.1 mg, 0.00750 mmol, 1.5 equiv)、ＯＭｅ−ＰＥＧ−ＮＨ₂ （MW：２，０００Ｄａ、Ｉｒｉｓ Ｂｉｏｔｅｃｈ社製）(10.4 mg, 0.00516 mmol)、およびDMT-MM (1.4 mg, 0.00506 mmol, 1.0 equiv) のTHF (1100 μＬ) およびMeOH (900 μＬ) 混合物を室温で7時間35分攪拌した。減圧下溶媒を留去した後、残渣にPEG-NH₂ (10.4 mg, 0.00516 mmol) およびDMT-MM (5.7 mg, 0.0206 mmol)、およびDMF (1 mL) を加えて室温で4時間38分攪拌した。DMT-MM (14.2 mg, 0.0513 mmol) を加えて1時間20分攪拌した。HPLCにて精製を行いBdev-13 (3.6 mg, 14.5%) を得た。m/z 1700付近を中心に22マス間隔で2価イオン群、m/z 1150付近を中心に15マス間隔で3価イオン群、m/z 900付近を中心に11マス間隔で4価イオン群が検出した。
（実施例５）Bdev-2, -4, -8,および-10合成のための中間体（化合物２５）の合成
Bdev-2,-4,-8,および-10合成の中間体の合成の概略図を以下に示す。

Bdev-5 (10.1 mg, 0.00750 mmol, 1.5 equiv), OMe-PEG-NH ₂ (MW: 2,000 Da, manufactured by Iris Biotech) (10.4 mg, 0.00516 mmol), and DMT-MM (1.4 mg, 0.00506 mmol) , 1.0 equiv) of THF (1100 μL) and MeOH (900 μL) were stirred at room temperature for 7 hours and 35 minutes. After evaporating the solvent under reduced pressure, PEG-NH ₂ (10.4 mg, 0.00516 mmol), DMT-MM (5.7 mg, 0.0206 mmol), and DMF (1 mL) were added to the residue, and the mixture was stirred at room temperature for 4 hours and 38 minutes. did. DMT-MM (14.2 mg, 0.0513 mmol) was added and stirred for 1 hour and 20 minutes. Purification by HPLC gave Bdev-13 (3.6 mg, 14.5%). Divalent ion group at 22 mass intervals around m / z 1700, trivalent ion group at 15 mass intervals around m / z 1150, and tetravalent ion group at 11 mass intervals around m / z 900 Detected.
Example 5 Synthesis of Intermediate (Compound 25) for Synthesis of Bdev-2, -4, -8, and -10
A schematic diagram of the synthesis of intermediates for Bdev-2, -4, -8, and -10 synthesis is shown below.

それぞれの合成は、以下のようにして行った。
化合物23の合成

Each synthesis was performed as follows.
Synthesis of compound 23

化合物13 (890 mg, 0.372 mmol)、化合物40 (776 mg, 0.559 mmol, 1.5 equiv)、およびPPh₃ (147 mg, 0.559 mmol, 1.5 equiv) をTHF (8 mL) に溶解し、DEAD (253 μＬ, 0.558 mmol, 1.5 equiv.) を加えて室温で54分間攪拌した。次に、PPh₃ (148 mg, 0.564 mmol, 1.5 equiv.) を加えて20分間攪拌した。次に、DEAD (84.4 μＬ, 0.186 mmol, 0.5 equiv.) を加えて2時間20分攪拌した。次に、PPh₃ (147 mg, 0.560 mmol, 1.5 equiv) を加えて14分間攪拌した。さらに、DEAD (33.7 μＬ, 0.0743 mmol, 0.2 equiv) を加えて34分間攪拌した。次に、PPh₃ (44.8 mg, 0.171 mmol, 0.5 equiv) を加えて2分間攪拌した。さらに、DEAD (33.7 μＬ, 0.0743 mmol, 0.2 equiv) を加えて3時間15分攪拌した。最後に、DEAD (33.7 μＬ, 0.0743 mmol, 0.2 equiv) を加えて21分間攪拌した。化合物4の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (CH₂CH₂/THF, 100 : 0 - 90 : 10) に付し、光延反応物 (1.05 g, 75.0%) を得た。

Compound 13 (890 mg, 0.372 mmol), Compound 40 (776 mg, 0.559 mmol, 1.5 equiv), and PPh ₃ (147 mg, 0.559 mmol, 1.5 equiv) were dissolved in THF (8 mL) and DEAD (253 μL , 0.558 mmol, 1.5 equiv.) Was added and stirred at room temperature for 54 minutes. Next, PPh ₃ (148 mg, 0.564 mmol, 1.5 equiv.) Was added and stirred for 20 minutes. Next, DEAD (84.4 μL, 0.186 mmol, 0.5 equiv.) Was added and stirred for 2 hours and 20 minutes. Next, PPh ₃ (147 mg, 0.560 mmol, 1.5 equiv) was added and stirred for 14 minutes. Furthermore, DEAD (33.7 μL, 0.0743 mmol, 0.2 equiv) was added and stirred for 34 minutes. Next, PPh ₃ (44.8 mg, 0.171 mmol, 0.5 equiv) was added and stirred for 2 minutes. Furthermore, DEAD (33.7 μL, 0.0743 mmol, 0.2 equiv) was added and stirred for 3 hours and 15 minutes. Finally, DEAD (33.7 μL, 0.0743 mmol, 0.2 equiv) was added and stirred for 21 minutes. The residue obtained by post-treatment equivalent to the synthesis of compound 4 was subjected to silica gel column chromatography (CH ₂ CH ₂ / THF, 100: 0-90: 10), and Mitsunobu reaction product (1.05 g, 75.0%) was added. Obtained.

光延反応物 (637 mg, 0.169 mmol) をCH₂Cl₂ (16.9 mL) に溶解し、TFE (1690 μＬ) およびTFA (169 μＬ) を加えて室温で40分間攪拌した。沈殿物をセライトろ過し、減圧下溶媒を留去した。化合物4の合成と同等の後処理を行い得た残渣をシリカゲルカラムクロマトグラフィー (CHCl₃/EtOH, 100 : 0 - 95 : 5) に付し、化合物23 (294 mg, 58.1%) を得た。
化合物24の合成

Mitsunobu reaction product (637 mg, 0.169 mmol) was dissolved in CH ₂ Cl ₂ (16.9 mL), TFE (1690 μL) and TFA (169 μL) were added, and the mixture was stirred at room temperature for 40 minutes. The precipitate was filtered through celite, and the solvent was distilled off under reduced pressure. The residue obtained by performing post-treatment equivalent to the synthesis of Compound 4 was subjected to silica gel column chromatography (CHCl ₃ / EtOH, 100: 0 to 95: 5) to obtain Compound 23 (294 mg, 58.1%).
Synthesis of compound 24

化合物23 (294 mg, 0.0970 mmol)、HOAt (15.8 mg, 0.116 mmol, 1.2 equiv)、およびHATU (44.7 mg, 0.118 mmol, 1.2 equiv) をTHF (19.4 mL) に溶解し、DIPEA (84.5 μＬ, 0.485 mmol, 5.0 equiv) を加えて室温で3時間10分間攪拌した。化合物4の合成と同等の後処理を行い化合物24 (284 mg, 97.2%) を得た。

Compound 23 (294 mg, 0.0970 mmol), HOAt (15.8 mg, 0.116 mmol, 1.2 equiv), and HATU (44.7 mg, 0.118 mmol, 1.2 equiv) were dissolved in THF (19.4 mL) and DIPEA (84.5 μL, 0.485). mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 3 hours and 10 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 24 (284 mg, 97.2%) was obtained.

Synthesis of compound 25

化合物24 (284 mg, 0.0943 mmol) をTHF (943 μＬ) に溶解し、PhSH (29.0 μＬ, 0.282 mmol, 3.0 equiv) およびDBU (42.3 μＬ, 0.283 mmol, 3.0 equiv) を加えて室温で2時間30分間攪拌した。化合物16の合成と同等の後処理を行い、Bdev-2,-4,-8,および-10合成の中間体である化合物25 (255 mg, 95.8%) を得た。
（実施例６）Bdev-2, -4, -8,および-10の合成
（１）Bdev-8の合成
Bdev-8の合成は、以下のようにして行った。

Compound 24 (284 mg, 0.0943 mmol) was dissolved in THF (943 μL), and PhSH (29.0 μL, 0.282 mmol, 3.0 equiv) and DBU (42.3 μL, 0.283 mmol, 3.0 equiv) were added for 2 hours at room temperature 30 Stir for minutes. The post-process equivalent to the synthesis | combination of the compound 16 was performed, and the compound 25 (255 mg, 95.8%) which is an intermediate body of Bdev-2, -4, -8, and -10 synthesis | combination was obtained.
(Example 6) Synthesis of Bdev-2, -4, -8, and -10 (1) Synthesis of Bdev-8
Bdev-8 was synthesized as follows.

化合物25 (55.1 mg, 0.0181 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (1810 μＬ) を加えて室温で3時間攪拌した。Bdev-7の合成と同等の後処理およびHPLC精製を行いBdev-8 (12.0 mg, 52.6%) を得た。HRMS m/z [M + H]⁺: calcd for C₆₄H₈₅N₁₂O₁₅: 1261.6257, found 1261.4668.
（２）Bdev-4の合成
化合物26の合成

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (1810 μL) was added to compound 25 (55.1 mg, 0.0181 mmol) and stirred at room temperature for 3 hours. Post-treatment and HPLC purification equivalent to the synthesis of Bdev-7 were performed to obtain Bdev-8 (12.0 mg, 52.6%). HRMS m / z [M + H] ⁺ : calcd for C ₆₄ H ₈₅ N ₁₂ O ₁₅ : 1261.6257, found 1261.4668.
(2) Synthesis of Bdev-4 synthesis compound 26

化合物25 (62.6 mg, 0.0222 mmol)、ミリスチン酸 (7.7 mg, 0.0337 mmol, 1.5 equiv)、HOAt (4.5 mg, 0.0331 mmol, 1.5 equiv)、およびHATU (12.8 mg, 0.0337 mmol, 1.5 equiv) をTHF (444 μＬ) に溶解し、DIPEA (19.3 μＬ, 0.111 mmol, 5.0 equiv) を加えて室温で3時間攪拌した。化合物4の合成と同等の後処理を行い化合物26 (56.9 mg, 84.7%) を得た。
Bdev-4の合成

Compound 25 (62.6 mg, 0.0222 mmol), myristic acid (7.7 mg, 0.0337 mmol, 1.5 equiv), HOAt (4.5 mg, 0.0331 mmol, 1.5 equiv), and HATU (12.8 mg, 0.0337 mmol, 1.5 equiv) in THF ( 444 μL), DIPEA (19.3 μL, 0.111 mmol, 5.0 equiv) was added, and the mixture was stirred at room temperature for 3 hours. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 26 (56.9 mg, 84.7%) was obtained.
Synthesis of Bdev-4

化合物26 (56.9 mg, 0.0188 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (1514 μＬ) を加えて室温で5分間攪拌した。TIS (46.9 mL) を加えて3時間55分攪拌した。Bdev-7の合成と同等の後処理を行いBdev-4 (4.0 mg, 14.5%) を得た。HRMS m/z [M + H]⁺: calcd for C₇₈H₁₁₁N₁₂O₁₆: 1471.8241, found 1471.8190.
（３）Bdev-2の合成
化合物27の合成

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (1514 μL) was added to compound 26 (56.9 mg, 0.0188 mmol), and the mixture was stirred at room temperature for 5 minutes. TIS (46.9 mL) was added and stirred for 3 hours and 55 minutes. Bdev-4 (4.0 mg, 14.5%) was obtained by post-treatment equivalent to the synthesis of Bdev-7. HRMS m / z [M + H] ⁺ : calcd for C ₇₈ H ₁₁₁ N ₁₂ O ₁₆ : 1471.8241, found 1471.8190.
(3) Synthesis of Bdev-2 Synthesis Compound 27

化合物24 (204 mg, 0.0724 mmol) をCH₂Cl₂ (1448 μＬ) に溶解し、Et₃N (20.4 μＬ, 0.145 mmol, 2.0 equiv) およびAc₂O (13.7 μＬ, 0.145 mmol, 2.0 equiv) を加えて室温で50分間攪拌した。化合物4の合成と同等の後処理を行い化合物27 (200 mg, 96.3%) を得た。

Compound 24 (204 mg, 0.0724 mmol) was dissolved in CH ₂ Cl ₂ (1448 μL), and Et ₃ N (20.4 μL, 0.145 mmol, 2.0 equiv) and Ac ₂ O (13.7 μL, 0.145 mmol, 2.0 equiv) were dissolved. In addition, the mixture was stirred at room temperature for 50 minutes. The post-process equivalent to the synthesis | combination of the compound 4 was performed, and the compound 27 (200 mg, 96.3%) was obtained.

化合物27 (200 mg, 0.0697 mmol) にTFA/TIS/H₂O = 95 : 5 : 5 の溶液 (6970 μＬ) を加えて室温で3時間攪拌した。Bdev-7の合成と同等の後処理を行いBdev-2 (54.4 mg, 59.8%) を得た。HRMS m/z [M + H]⁺: calcd for C₆₆H₈₇N₁₂O₁₆: 1303.6363, found 1303.5524.であった。
（４）Bdev-10の合成 Bdev-2 synthesis

A solution of TFA / TIS / H ₂ O = 95: 5: 5 (6970 μL) was added to compound 27 (200 mg, 0.0697 mmol), and the mixture was stirred at room temperature for 3 hours. Bdev-2 (54.4 mg, 59.8%) was obtained by post-treatment equivalent to the synthesis of Bdev-7. HRMS m / z [M + H] ⁺ : calcd for C ₆₆ H ₈₇ N ₁₂ O ₁₆ : 1303.6363, found 1303.5524.
(4) Synthesis of Bdev-10

Bdev-2 (13.1 mg, 0.0101 mmol) およびSUNBRIGHT ME-020AS（日油株式会社製） (40.4 mg, 0.0202 mmol, 2.0 equiv) のDMF (404 μＬ) にEt₃N (2.84 μＬ, 0.0202 mmol, 2.0 equiv) を加えて室温で22時間40分攪拌した。HPLC精製しBdev-10 (16.8 mg, 48.9%) を得た。
m/z 1700付近を中心に22マス間隔で2価イオンが検出された。

Et ₃ N (2.84 μL, 0.0202 mmol, 2.0) in DMF (404 μL) of Bdev-2 (13.1 mg, 0.0101 mmol) and SUNBRIGHT ME-020AS (manufactured by NOF Corporation) (40.4 mg, 0.0202 mmol, 2.0 equiv) equiv) was added and the mixture was stirred at room temperature for 22 hours and 40 minutes. HPLC purification gave Bdev-10 (16.8 mg, 48.9%).
Divalent ions were detected at intervals of 22 squares around m / z 1700.

Comparative Example Synthesis of Comparative Example Compound As a comparative example, a WP9QY peptide (disulfide bridge: disulfide bridge: compound number: STD) was used. The WP9QY peptide is a process in which amino acids are sequentially condensed to 2,4 docosyloxybenzyl alcohol, a linear sequence is synthesized, a disulfide bond is formed by iodine oxidation, and 2,4 docosyloxybenzyl alcohol is cleaved with trifluoroacetic acid. It was prepared after.

(Reference Example) As a synthesis reference example of a reference example, a modified product in which the cross-linked moiety was modified to a thioether bridge (Compound No. Comp. 1) or an olefin bridge (Compound No. Comp. 3) was used. The thioether bridged WP9QY peptide was synthesized according to the synthetic scheme described in FIG. The olefin bridged WP9QY peptide was synthesized according to the synthesis scheme described in FIG.

Example 7 Resistance to Peptidase Carboxypeptidase and chymotrypsin were used to examine the resistance of the peptide of the present invention to peptidase. Resistance to peptidase was measured as follows.
(Decomposition by carboxypeptidase)
An enzyme solution of carboxypeptidase purchased from SIGMA was prepared at 1 U / ml in PBS and incubated at 37 ° C. in a water bath. Next, the peptide volume adjusted to 5 mg / ml with DMSO: pure water = 1: 1 mixed solvent is adjusted to 1 mg / ml with PBS (-), the enzyme solution and the peptide solution are mixed at 4: 1, and the peptide is mixed. The final concentration of was 0.2 mg / ml. Thereafter, the reaction was carried out immediately at 37 ° C. 0.1 ml was sampled over time, and the reaction was stopped by adding a reaction stop solution (25% TFA in acetonitrile 20 μL). Each sample was measured by HPLC, and peptide degradation by peptidase was measured. Sampling timing is 0 min, 0.5 min, 1 min, 2 min. If degradation of 50% or more is not obtained even in 2 min, 0.5 hr later, 1 hr later, 3 hr Thereafter, in addition, it was carried out after 6 hours in some cases, and for a peptide that was stable for a long time, sampling was performed as necessary in the range from 24 hours to 168 hours later. The time for half of the added peptide to degrade was measured. The results are shown in Table 3.

(Degradation by chymotrypsin)
An enzyme solution of chymotrypsin purchased from SIGMA was adjusted to 4 U / ml with 0.1 M Tris-HCl (pH 8.0), and kept at 37 ° C. in a water bath. Next, a peptide solution prepared at 5 mg / ml in DMSO: pure water = 1: 1 was prepared at 1 mg / ml in 0.1 M Tris-HCl (pH 8.0), and the enzyme solution and the peptide solution were mixed at 4: 1. The mixture was mixed so that the final peptide concentration was 0.2 mg / ml. Thereafter, the reaction was carried out immediately at 37 ° C. 0.1 ml was sampled over time, and the reaction was stopped by adding a reaction stop solution (25% TFA in acetonitrile 20 μL). Each sample was measured by HPLC, and peptide degradation by chymotrypsin was measured. Sampling timing is 0 min, 0.5 min, 1 min, 2 min. If degradation of 50% or more cannot be obtained in 2 min, further 0.5 hr, 1 hr, 3 hr later The time taken for half of the added peptide to degrade was measured. The results are shown in Table 3.

  The above results indicate that the WP9QY peptidomimetic of the present invention prepared according to the crosslinking method of the present invention has improved degradation resistance to peptidase compared to the WP9QY peptide peptide of disulfide bridge or thioether bridge. It was.

(Example 8: Osteoclast differentiation inhibition test)
The osteoclast differentiation inhibitory activity of the WP9QY peptide mimetic of the present invention was examined using cultured osteoclasts as follows.
Bone marrow cells were collected from self-bred C57BL / 6jjc1 mice (weight 18-20 g), and RANKL (50 ng / ml, recombinant human RANKL purchased from Wako Pure Chemical Industries) and M-CSF (25 ng / ml, recombinant) In the presence of each WP9QY cross-linked peptidomimetic in medium containing α-MEM, 10% FBS (SIGMA), penicillin 100U, streptomycin 100 μg / ml, containing human M-CSF purchased from R & D System Cultured for 4 days.

Each peptide was prepared at 5 mg / ml in DMSO: MQ = 1: 1 (used in the experiment shown in FIG. 5 below) or 4 mM in DMSO: MQ = 1: 9 (used in the experiment shown in FIG. 6 below). The solution was diluted with a culture medium and sterilized by filtration with a filter (0.22 μm) to obtain a peptide preparation. Each peptide preparation solution was added so that it might become the target final concentration, and the bone peritoneal cell was cultured on the said conditions.
The culture medium was replaced with a new medium on the third day of culture, and fixed with 3.6% formalin (36% formalin diluted with PBS (-)) on the fourth day, and then acetone: ethanol = 1: 1. Permeabilized for 30 seconds, dried for 20 minutes, and stained with TRAP (tartrate-resistant acid phosphatase) to confirm mature osteoclasts. The peptide concentration was cultured such that the final concentration in the medium was 0.3, 1, 3, 10 μM. As controls, no peptide addition and WP9QY peptide (disulfide bond) addition were used. The results are shown in FIG. 5 and FIG. It was shown that Bdev-2 and Bdev-3 have a stronger osteoclast differentiation inhibitory effect than WP9QY peptide (disulfide bond) at low concentrations.

Claims (5)

An osteoclast osteoclast growth inhibitor comprising a peptide represented by the following chemical formula:

Wherein Z ₁ and Z ₃ are each independently a hydrogen atom, an unsubstituted or substituted acyl group having 1 to 8 carbon atoms, an unsubstituted or substituted alkyl group having 1 to 8 carbon atoms, and —C Selected from the group consisting of polyethylene glycol having a molecular weight of 100 to 10,000 Da represented by (═O) —CH ₂ CH ₂ (OCH ₂ CH ₂ ) nOCH ₂ CH ₂ OCH ₃ (n is an integer), Z ₂ hydroxyl group, an amino group, an unsubstituted or substituted mono-alkylamino group having 1 to 8 carbon atoms, molecular weight 100, represented by _{-NH-CH 2 CH 2 (OCH} 2 CH 2) nOCH 2 CH 2 OCH 3 (Selected from the group consisting of 10,000 Da polyethylene glycol) The inhibitor according to claim 1, selected from the group consisting of peptides represented by the following chemical formulas:

,and

(Here, Ac represents an acetyl group, and polyethylene glycol represents polyethylene glycol having a number average molecular weight of 500 to 2000 Da).   A pharmaceutical composition comprising the inhibitor according to claim 1 or 2 as an active ingredient.   It is a therapeutic agent for diseases selected from the group consisting of osteoporosis, Paget's disease, rheumatoid arthritis and other forms of inflammatory arthritis, osteoarthritis, prosthetic breakage, osteolytic tumors, myeloma and tumor metastasis to bone The pharmaceutical composition according to claim 3.   A method of treating a disease selected from the group consisting of osteoporosis, Paget's disease, rheumatoid arthritis and other forms of inflammatory arthritis, osteoarthritis, prosthetic breakage, osteolytic tumors, myeloma and tumor metastasis to bone. A method of treatment comprising administering to a mammal in need of such treatment at least one of the peptides of claim 1 or 2 in a therapeutically effective amount.

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