JP2017043615A - Cyclopropane amino acid unit that improves membrane permeability and/or metabolic stability of cyclic peptide - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a cyclopropane amino acid unit that improves the membrane permeability and/or metabolic stability of a cyclic peptide.SOLUTION: The present invention provides a method whereby any cyclopropane amino acid unit represented by formula (I-i') to formula (IV-viii') is selected, the cyclopropane amino acid unit is introduced into the ring of a cyclic peptide, and the membrane permeability and/or metabolic stability of the cyclic peptide is improved.SELECTED DRAWING: None

Description

  The present invention relates to cyclopropane amino acid units that improve the membrane permeability and / or metabolic stability of cyclic peptides.

Cyclic peptides are useful as pharmaceuticals because they can act on drug targets that are difficult to control with small molecules such as intracellular protein-protein interactions and protein-nucleic acid interactions. Several cyclic peptides derived from natural products that permeate cell membranes have been found (Non-patent Document 1), but it is still difficult to artificially design and synthesize cyclic peptides with high membrane permeability. So far, there has been a report on the relationship between the conformation of the entire molecule of the cyclic peptide and the membrane permeability (Non-Patent Documents 2 to 4), and a technique for rationally controlling the conformation is required. In addition, peptides are generally known to be easily degraded by in vivo metabolic enzymes such as proteases, and since it is not easy to develop them as pharmaceuticals, a technique for overcoming them is also required.
On the other hand, the cyclopropane ring is the smallest carbocyclic ring and has no steric freedom, and the steric and stereoelectronic effects such as cyclopropane strain and bipartite conformation can control the molecular conformation over a wide range. It is possible (Non-Patent Documents 5 and 6). Various bioactive compounds have been designed and synthesized by utilizing such properties of the cyclopropane ring. For example, Non-Patent Documents 7 and 8 disclose compounds having a cyclopropane ring as an antagonist of histamine H3 and / or H4 receptor. Non-Patent Document 9 discloses an amino acid having a cyclopropane ring as an agonist compound of GABA _C receptor. Non-Patent Document 10 discloses a method for synthesizing an amino acid having a cyclopropane ring. Non-Patent Document 11 discloses a compound having a steroid-spiroketal-oligopeptide structure having a cyclopropane ring, and the oligopeptide structure includes an amino acid having a cyclopropane ring. However, all of these examples are introduced into low molecules, and there is no example in which an amino acid having a cyclopropane ring is introduced into a cyclic peptide to realize conformational control.

Curr. Top. Med. Chem. 2013, 13, 821-836 J. Am. Chem. Soc. 2006, 128, 2510-2511 Nat. Chem. Biol. 2011, 7, 810-817 ACS Med. Chem. Lett. 2014, 5, 1148-1151 J. Org. Chem. 1996, 61, 915-923 J. Med. Chem. 1996, 39, 4844-4852 J. Org. Chem. 2002, 67, 1669-1677 J. Med. Chem. 2006, 49, 5587-5596 Tetrahedron Asymmetry 1998, 9, 2533-2548 Organic Letters 2005, 7 (1), 103-106 Organic Letters 2009, 11 (1), 65-68

  An object of the present invention is to provide a cyclopropane amino acid unit that improves the membrane permeability and / or metabolic stability of a cyclic peptide.

（式中、
Ｒ^１、Ｒ^２、Ｒ^３、およびＲ^４は、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５Ａは、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５およびＲ^６は、それぞれ独立して、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基である）
で示されるいずれかのシクロプロパンアミノ酸ユニットを選択し、環状ペプチドの環中に当該シクロプロパンアミノ酸ユニットを導入し、当該環状ペプチドの膜透過性および／または代謝安定性を改善する方法。 The present invention relates to the following.
[1] The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbon A cyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group)
A method for improving the membrane permeability and / or metabolic stability of the cyclic peptide by selecting any of the cyclopropane amino acid units represented by the above formula and introducing the cyclopropane amino acid unit into the ring of the cyclic peptide.

（式中の記号は前記[１]と同意義）
で示されるいずれかのシクロプロパンアミノ酸ユニットを、環状ペプチドの環中に導入し、当該シクロプロパンアミノ酸ユニットを導入する前の環状ペプチドと膜透過性および／または代謝安定性を比較し、当該環状ペプチドの膜透過性および／または代謝安定性を改善するシクロプロパンアミノ酸ユニットを選択する方法。 [2] The following formula:

(The symbols in the formula are as defined in [1] above)
Is introduced into the ring of the cyclic peptide, and the cyclic peptide before introduction of the cyclopropane amino acid unit is compared with membrane permeability and / or metabolic stability. For selecting a cyclopropane amino acid unit that improves the membrane permeability and / or metabolic stability of.

（式中、
Ｒ^１、Ｒ^２、Ｒ^３、およびＲ^４は、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、または置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５Ａは、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５およびＲ^６は、それぞれ独立して、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｘ^Ａは、Ｆｍｏｃ、Ａｌｌｏｃ、およびＴｒｏｃから選択される一の保護基であり、
Ｘは、水素またはＢｏｃ、Ｃｂｚ、Ｆｍｏｃ、Ａｌｌｏｃ、およびＴｒｏｃから選択される一の保護基であり、
Ｙは、水素、置換もしくは非置換のアルキル、または核酸である）
で示されるいずれかの化合物（ただし、以下の化合物を除く

）。 [3] The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic A carbocyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
X ^A is one protecting group selected from Fmoc, Alloc, and Troc;
X is hydrogen or one protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc;
Y is hydrogen, substituted or unsubstituted alkyl, or nucleic acid)
Any of the compounds shown below (excluding the following compounds)

).

[4] Any compound represented by formulas (II-i) to (II-viii), (III-i) to (III-viii), and (IV-i) to (IV-viii), The compound according to [3], wherein X is hydrogen.

[5] Any compound represented by formulas (II-i) to (II-viii), (III-i) to (III-viii), and (IV-i) to (IV-viii), , X is one protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc, [3].

[6] The compound according to any one of [3] to [5], wherein Y is hydrogen.

（式中の記号は前記[１]と同意義である）
で示されるいずれかの基を環中に有する環状ペプチド。 [7] The following formula:

(The symbols in the formula are as defined in [1] above)
The cyclic peptide which has either group shown by in a ring.

で示されるいずれかの基を環中に有する環状ペプチド。 [8] The following formula:

The cyclic peptide which has either group shown by in a ring.

By introducing the cyclopropane amino acid unit according to the present invention into a cyclic peptide, the three-dimensional structure of the cyclic peptide can be changed, and the membrane permeability and / or metabolic stability of the cyclic peptide can be improved. Cyclic peptides containing such cyclopropane amino acid units are useful as pharmaceuticals. In another aspect, a library of cyclic peptides having cyclopropane amino acid units is also useful as a screening source for searching for pharmaceutical lead compounds.
The cyclopropane amino acid unit according to the present invention has various three-dimensional structures, and when the cyclopropane amino acid unit is introduced into the cyclic peptide, the three-dimensional structure of the cyclic peptide is specifically determined depending on each cyclopropane amino acid unit. It is believed that immobilization and structural stability can improve membrane permeability and / or metabolic stability.
Furthermore, in a cyclic peptide having a specific activity, the cyclopropane amino acid unit according to the present invention is introduced into an amino acid site that is assumed not to affect the activity, so that the cyclic peptide is not affected. It may be possible to improve the membrane permeability and / or metabolic stability.

The meaning of each term used in this specification will be described below. Unless otherwise specified, each term is used in the same meaning when used alone or in combination with other terms.
The term “consisting of” means having only the configuration requirements.
The term “comprising” is not limited to the constituent elements and means that elements not described are not excluded.

  As a specific method of using the cyclopropane amino acid unit of the present invention, for example, a cyclic peptide having a known sequence has a specific activity (function), but the cyclic peptide has a membrane permeability and / or metabolism. When it is difficult to apply to pharmaceuticals due to poor stability, the cyclopropane amino acid unit of the present invention can be introduced to improve the membrane permeability and / or metabolic stability. As a specific method, in order to identify amino acids located at sites that do not affect the activity of the cyclic peptide, each amino acid constituting the cyclic peptide is replaced with a small amino acid such as alanine, and the activity is increased. If maintained, the amino acid located at the substituted site can be identified as an amino acid that does not affect activity. Next, an amino acid that does not affect the activity is substituted with the cyclopropane amino acid unit of the present invention, and studies are made to improve the membrane permeability and / or metabolic stability of the cyclic peptide. At this time, since it is unclear which cyclopropane amino acid unit is most preferable, cyclopropane amino acid units having various three-dimensional structures represented by the formulas (Ii ′) to (IV-viii ′) are introduced. The present invention searches for cyclopropane amino acid units that improve the membrane permeability and / or metabolic stability of the cyclic peptide while maintaining the activity. In particular, when improving membrane permeability, the fat solubility of the cyclic peptide is important. For example, the fat solubility derived from the amino acid composition of the cyclic peptide, the fat solubility of the amino acid substituted for the cyclopropane amino acid unit, and the cyclopropane amino acid It is believed that the fat solubility of the side chain attached to the carbon adjacent to the unit cyclopropane is important. Therefore, it is considered preferable to design so that the fat solubility does not decrease with reference to values such as ClogP. By introducing the cyclopropane amino acid unit designed in this way into a site that does not affect the activity of the cyclic peptide, a cyclic peptide having improved membrane permeability and / or metabolic stability while maintaining the activity can be obtained. Can be obtained.

“Alkyl” includes straight or branched hydrocarbon groups having 1 to 15 carbon atoms, preferably 1 to 10 carbon atoms, more preferably 1 to 6 carbon atoms, and still more preferably 1 to 4 carbon atoms. To do. For example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, isohexyl, n-heptyl, isoheptyl, n-octyl , Isooctyl, n-nonyl, n-decyl and the like.
Preferable embodiments of “alkyl” include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl and n-pentyl. Further preferred examples include methyl, ethyl, n-propyl, isopropyl and tert-butyl.

“Alkenyl” has 2 to 15 carbon atoms, preferably 2 to 10 carbon atoms, more preferably 2 to 6 carbon atoms, and further preferably 2 to 4 carbon atoms, having one or more double bonds at any position. These linear or branched hydrocarbon groups are included. For example, vinyl, allyl, propenyl, isopropenyl, butenyl, isobutenyl, prenyl, butadienyl, pentenyl, isopentenyl, pentadienyl, hexenyl, isohexenyl, hexadienyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, decenyl, tridecenyl, decenyl Etc.
Preferred embodiments of “alkenyl” include vinyl, allyl, propenyl, isopropenyl and butenyl.

“Alkynyl” has 2 to 10 carbon atoms, preferably 2 to 8 carbon atoms, more preferably 2 to 6 carbon atoms, more preferably 2 to 4 carbon atoms, having one or more triple bonds at any position. Includes straight chain or branched hydrocarbon groups. Furthermore, you may have a double bond in arbitrary positions. Examples include ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl and the like.
Preferred embodiments of “alkynyl” include ethynyl, propynyl, butynyl and pentynyl.

The “aromatic carbocyclic group” means a cyclic aromatic hydrocarbon group having one or more rings. For example, phenyl, naphthyl, anthryl, phenanthryl and the like can be mentioned.
A preferred embodiment of the “aromatic carbocyclic group” includes phenyl.

単環の非芳香族炭素環式基としては、炭素数３〜１６が好ましく、より好ましくは炭素数３〜１２、さらに好ましくは炭素数４〜８である。例えば、シクロプロピル、シクロブチル、シクロペンチル、シクロヘキシル、シクロヘプチル、シクロオクチル、シクロノニル、シクロデシル、シクロプロペニル、シクロブテニル、シクロペンテニル、シクロヘキセニル、シクロヘプテニル、シクロヘキサジエニル等が挙げられる。
２環以上の非芳香族炭素環式基としては、炭素数８〜２０が好ましく、より好ましくは炭素数８〜１６である。例えば、インダニル、インデニル、アセナフチル、テトラヒドロナフチル、フルオレニル等が挙げられる。 The “non-aromatic carbocyclic group” means a cyclic saturated hydrocarbon group or a cyclic non-aromatic unsaturated hydrocarbon group having one or more rings. The “non-aromatic carbocyclic group” having two or more rings includes those obtained by condensing a ring in the above “aromatic carbocyclic group” to a monocyclic or two or more non-aromatic carbocyclic groups.
Furthermore, the “non-aromatic carbocyclic group” includes a group that forms a bridge or a spiro ring as described below.

The monocyclic non-aromatic carbocyclic group preferably has 3 to 16 carbon atoms, more preferably 3 to 12 carbon atoms, and still more preferably 4 to 8 carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclohexadienyl, and the like.
The non-aromatic carbocyclic group having 2 or more rings preferably has 8 to 20 carbon atoms, and more preferably has 8 to 16 carbon atoms. For example, indanyl, indenyl, acenaphthyl, tetrahydronaphthyl, fluorenyl and the like can be mentioned.

“Aromatic heterocyclic group” means a monocyclic or bicyclic or more aromatic cyclic group having one or more heteroatoms arbitrarily selected from O, S and N in the ring To do. The aromatic heterocyclic group having two or more rings includes those obtained by condensing a ring in the above “aromatic carbocyclic group” to a monocyclic or two or more aromatic heterocyclic group.
The monocyclic aromatic heterocyclic group is preferably 5 to 8 members, more preferably 5 or 6 members. Examples include pyrrolyl, imidazolyl, pyrazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazolyl, triazinyl, tetrazolyl, furyl, thienyl, isoxazolyl, oxazolyl, oxadiazolyl, isothiazolyl, thiazolyl, thiadiazolyl and the like.
The bicyclic aromatic heterocyclic group is preferably 8 to 10 members, more preferably 9 or 10 members. For example, indolyl, isoindolyl, indazolyl, indolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, naphthyridinyl, quinoxalinyl, purinyl, pteridinyl, benzimidazolyl, benzisoxazolyl, benzoxazoazolyl, benzoxadiazolyl, benzoisodiazolyl Ril, benzothiazolyl, benzothiadiazolyl, benzofuryl, isobenzofuryl, benzothienyl, benzotriazolyl, imidazopyridyl, triazolopyridyl, imidazothiazolyl, pyrazinopyridazinyl, oxazolopyridyl, thiazolopyridyl, etc. Is mentioned.
Examples of the aromatic heterocyclic group having 3 or more rings include carbazolyl, acridinyl, xanthenyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, dibenzofuryl and the like.

単環の非芳香族複素環式基としては、３〜８員が好ましく、より好ましくは５員または６員である。例えば、ジオキサニル、チイラニル、オキシラニル、オキセタニル、オキサチオラニル、アゼチジニル、チアニル、チアゾリジニル、ピロリジニル、ピロリニル、イミダゾリジニル、イミダゾリニル、ピラゾリジニル、ピラゾリニル、ピペリジル、ピペラジニル、モルホリニル、モルホリノ、チオモルホリニル、チオモルホリノ、ジヒドロピリジル、テトラヒドロピリジル、テトラヒドロフリル、テトラヒドロピラニル、ジヒドロチアゾリル、テトラヒドロイソチアゾリル、ジヒドロオキサジニル、ヘキサヒドロアゼピニル、テトラヒドロジアゼピニル、テトラヒドロピリダジニル、ヘキサヒドロピリミジニル、ジオキソラニル、ジオキサジニル、アジリジニル、ジオキソリル、オキセパニル、チオラニル、チイニル、チアジニル等が挙げられる。
２環以上の非芳香族複素環式基としては、８〜２０員が好ましく、より好ましくは８〜１０員である。例えば、インドリニル、イソインドリニル、クロマニル、イソクロマニル等が挙げられる。 “Non-aromatic heterocyclic group” means a monocyclic or bicyclic or more cyclic non-aromatic cyclic group having at least one hetero atom selected from O, S and N in the ring. Means group. The non-aromatic heterocyclic group having 2 or more rings is a monocyclic or 2 or more non-aromatic heterocyclic group, the above “aromatic carbocyclic group”, “non-aromatic carbocyclic group”, and / Or each condensed ring in the “aromatic heterocyclic group” is condensed, and further, the ring in the above “aromatic heterocyclic group” is condensed to a monocyclic or two or more non-aromatic carbocyclic groups. Also included.
Furthermore, the “non-aromatic heterocyclic group” includes a group that forms a bridge or a spiro ring as described below.

The monocyclic non-aromatic heterocyclic group is preferably 3 to 8 members, more preferably 5 or 6 members. For example, dioxanyl, thiranyl, oxiranyl, oxetanyl, oxathiolanyl, azetidinyl, thianyl, thiazolidinyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, morpholinyl, morpholino, thiomorpholinyl, morpholino, thiomorpholinyl, morpholino, thiomorpholinyl Furyl, tetrahydropyranyl, dihydrothiazolyl, tetrahydroisothiazolyl, dihydrooxazinyl, hexahydroazepinyl, tetrahydrodiazepinyl, tetrahydropyridazinyl, hexahydropyrimidinyl, dioxolanyl, dioxazinyl, aziridinyl, dioxolyl, Oxepanyl, thiolanyl, thiinyl, thiazinyl, etc. That.
The non-aromatic heterocyclic group having two or more rings is preferably 8 to 20 members, more preferably 8 to 10 members. Examples thereof include indolinyl, isoindolinyl, chromanyl, isochromanyl and the like.

“Boc” refers to a tert-butoxycarbonyl group which is a kind of protecting group.
“Cbz” refers to a benzyloxycarbonyl group which is a kind of protecting group.
“Fmoc” refers to a 9-fluorenylmethyloxycarbonyl group which is a kind of protecting group.
“Alloc” refers to an allyloxycarbonyl group which is a kind of protecting group.
“Troc” refers to a 2,2,2-trichloroethoxycarbonyl group which is a kind of protecting group.

  “Nucleic acid” refers to a polymer of nucleotides of any length. The sequence is not particularly limited, but tRNA is preferred. In the present invention, by adding tRNA to the carboxyl group of the cyclopropane amino acid unit, the cyclopropane amino acid unit can be introduced into the peptide using a biotechnology technique.

“Peptide” refers to a polymer of amino acids of any length. Includes linear, cyclic, and branched ones. Here, the amino acids include alpha amino acids, beta amino acids, gamma amino acids, and delta amino acids, and preferably include alpha amino acids. Alpha amino acids include D-form and L-form, and preferably L-form. Amino acids include not only naturally occurring amino acids but also synthesized amino acids. Preferred are naturally occurring amino acids, preferably essential amino acids. Aspartic acid, glutamic acid, lysine, arginine, histidine, glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, proline, phenylalanine, tyrosine, tryptophan and the like are preferable. The amino acids of the present invention also include amino acids that are made by modification after protein synthesis. For example, cystine, hydroxyproline, hydroxylysine, thyroxine, O-phosphoserine, desmosine and the like.
“Cyclic peptide” refers to a peptide in which a cyclic structure is formed by an amide bond between a nitrogen atom at the N-terminus of the peptide and a carbonyl carbon at the C-terminus. In this invention, the number of the amino acids which comprise a preferable cyclic peptide is 3-15, More preferably, it is 4-10, More preferably, it is 5-8, Most preferably, it is 6.

（式中、
Ｒ^１、Ｒ^２、Ｒ^３、およびＲ^４は、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５Ａは、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５およびＲ^６は、それぞれ独立して、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基である）
で示される基を意味する。 A “cyclopropane amino acid unit” is an unnatural amino acid having an amino group via a methylene bonded to a carbon on the cyclopropane ring, and further a carbon bonded to the methylene on the cyclopropane ring. And a structure having a carboxyl group on another carbon via a single bond or methylene, and having one of the following structures. Note that the cyclopropane ring ^may have a substituent group, as indicated by R ^1, R 2, ^{R 3} or ^{R 4,.} The methylene may have a substituent as represented by R ^5A , R ⁵ , or R ⁶ of the present invention.
Specifically, the following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbon A cyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group)
Means a group represented by

（式中、
Ｒ^１、Ｒ^２、Ｒ^３、およびＲ^４は、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、または置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５Ａは、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５およびＲ^６は、それぞれ独立して、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｘ^Ａは、Ｆｍｏｃ、Ａｌｌｏｃ、およびＴｒｏｃから選択される一の保護基であり、
Ｘは、水素またはＢｏｃ、Ｃｂｚ、Ｆｍｏｃ、Ａｌｌｏｃ、およびＴｒｏｃから選択される一の保護基であり、
Ｙは、水素、置換もしくは非置換のアルキル、または核酸である）
で示されるいずれかの化合物。 When a cyclopropane amino acid unit is introduced into a cyclic peptide and / or when a cyclopropane amino acid unit is synthesized, a compound represented by the following formula is used.
The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic A carbocyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
X ^A is one protecting group selected from Fmoc, Alloc, and Troc;
X is hydrogen or one protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc;
Y is hydrogen, substituted or unsubstituted alkyl, or nucleic acid)
Any compound represented by

（式中、
Ｒ^１、Ｒ^２、Ｒ^３、およびＲ^４は、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５Ａは、それぞれ独立して、水素、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基であり、
Ｒ^５およびＲ^６は、それぞれ独立して、置換もしくは非置換のアルキル、置換もしくは非置換のアルケニル、置換もしくは非置換のアルキニル、置換もしくは非置換の芳香族炭素環式基、置換もしくは非置換の非芳香族炭素環式基、置換もしくは非置換の芳香族複素環式基、または置換もしくは非置換の非芳香族複素環式基である）
で示される基からなる群から選択されるいずれかの基を環中に有する環状ペプチドは、当該基を導入する前の環状ペプチドと比較して、膜透過性および／または代謝安定性が改善され得る。特に、好ましくは、以下の式で示される基からなる群から選択されるいずれかの基を環中に有する環状ペプチドは当該基を導入する前の環状ペプチドと比較して、膜透過性および／または代謝安定性が改善され得る。
以下の式：

The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbon A cyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group)
The cyclic peptide having any group selected from the group consisting of the groups shown in the above in the ring has improved membrane permeability and / or metabolic stability compared to the cyclic peptide before the introduction of the group. obtain. In particular, preferably, the cyclic peptide having any group selected from the group consisting of the groups represented by the following formulas in the ring is more membrane permeable and / or less than the cyclic peptide before the group is introduced. Alternatively, metabolic stability can be improved.
The following formula:

“Introduction” means that one or more adjacent amino acids in a peptide sequence are replaced with one cyclopropane amino acid unit, and that one or more cyclopropane amino acid units are inserted between adjacent amino acids in the peptide sequence. To be done. Furthermore, cyclopropane amino acid units may be introduced at a plurality of positions in the peptide sequence. The number of cyclopropane amino acid units introduced into one cyclic peptide may be one, or two or more. In particular, the preferred number of cyclopropane amino acid units introduced into one cyclic peptide is one.
The case where a plurality of adjacent amino acids and one cyclopropane amino acid unit in the peptide sequence are replaced includes the case where two or three adjacent amino acids and one cyclopropane amino acid unit are replaced.

  The present invention is a method for improving the membrane permeability and / or metabolic stability of a cyclic peptide by introducing a cyclopropane amino acid unit selected from the cyclopropane amino acid units of the present invention into the ring of the cyclic peptide. In another embodiment, a cyclopropane amino acid unit is introduced into the ring of the cyclic peptide, and the membrane permeability and / or metabolic stability is compared with the cyclic peptide before the introduction of the cyclopropane amino acid unit. It is a method of selecting a cyclopropane amino acid unit that improves the membrane permeability and / or metabolic stability of a cyclic peptide.

As a characteristic of cyclic peptides with poor membrane permeability, for example, there are many highly polar amino acids (for example, aspartic acid, glutamic acid, arginine, histidine, lysine, asparagine, serine, glutamine, threonine, tyrosine, cysteine, etc.) in the cyclic peptide. Examples include cyclic peptides (for example, cyclic peptides in which highly polar amino acids account for 10% or more) and cyclic peptides in which an amide bond that is a binding site between amino acids and amino acids is exposed to the outside of the cyclic peptide.
As a characteristic of a cyclic peptide having poor metabolic stability, for example, a cyclic peptide having an amino acid sequence recognized by a peptidase or a protease can be mentioned. In addition, in the case of a cyclic peptide having high fat solubility, it can be degraded in vivo by metabolic enzymes such as microsomes.

Although the amino acid of the cyclic peptide which substitutes a cyclopropane amino acid unit is not specifically limited, The amino acid which is comparable hydrophobicity (For example, the value of ClogP is comparable) is preferable.
The amino acids at both ends when the cyclopropane amino acid unit is inserted are not particularly limited.

Examples of the method for confirming the membrane permeability of the cyclic peptide include: For example, the membrane permeability can be confirmed according to the membrane permeability test described in Test Example 3 of the present specification.
Examples of the method for confirming the metabolic stability of the cyclic peptide include a serum stability test, a trypsin stability test, and a protease stability test. For example, metabolic stability can be confirmed according to the metabolic stability test described in Test Example 1 of the present specification.

  When the membrane permeability and / or metabolic stability of the cyclic peptide is compared with the cyclic peptide before introduction of the cyclopropane amino acid unit, and the membrane permeability and / or metabolic stability of the cyclic peptide is improved, the cyclopropane amino acid unit Can be selected as a cyclopropane amino acid unit that improves the membrane permeability and / or metabolic stability of the cyclic peptide.

Examples of the substituent of “substituted or unsubstituted alkyl”, “substituted or unsubstituted alkenyl”, and “substituted or unsubstituted alkynyl” include the following substituents. The carbon atom at any position may be bonded to one or more groups selected from the following substituents.
Substituents: halogen, hydroxy, carboxy, amino, imino, hydroxyamino, hydroxyimino, formyl, formyloxy, carbamoyl, sulfamoyl, sulfanyl, sulfino, sulfo, thioformyl, thiocarboxy, dithiocarboxy, thiocarbamoyl, cyano, nitro, nitroso , Azide, hydrazino, ureido, amidino, guanidino, pentafluorothio, trialkylsilyl, alkyloxy, alkenyloxy, alkynyloxy, haloalkyloxy, alkylcarbonyl, alkenylcarbonyl, alkynylcarbonyl, monoalkylamino, dialkylamino, alkylsulfonyl, Alkenylsulfonyl, alkynylsulfonyl, monoalkylcarbonylamino, dialkylcarbonylamino, Alkylsulfonylamino, dialkylsulfonylamino, alkylimino, alkenylimino, alkynylimino, alkylcarbonylimino, alkenylcarbonylimino, alkynylcarbonylimino, alkyloxyimino, alkenyloxyimino, alkynyloxyimino, alkylcarbonyloxy, alkenylcarbonyloxy, alkynyl Carbonyloxy, alkyloxycarbonyl, alkenyloxycarbonyl, alkynyloxycarbonyl, alkylsulfanyl, alkenylsulfanyl, alkynylsulfanyl, alkylsulfinyl, alkenylsulfinyl, alkynylsulfinyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted Non-aromatic carbocyclic group, substituted or unsubstituted aroma Heterocyclic group, and a substituted or unsubstituted non-aromatic heterocyclic group.

“Substituted or unsubstituted aromatic carbocyclic group”, “Substituted or unsubstituted nonaromatic carbocyclic group”, “Substituted or unsubstituted aromatic heterocyclic group”, and “Substituted or unsubstituted aromatic carbocyclic group” Examples of the substituent on the ring of the “non-aromatic heterocyclic group” include the following substituents. An atom at any position on the ring may be bonded to one or more groups selected from the following substituents.
Substituents: halogen, hydroxy, carboxy, amino, imino, hydroxyamino, hydroxyimino, formyl, formyloxy, carbamoyl, sulfamoyl, sulfanyl, sulfino, sulfo, thioformyl, thiocarboxy, dithiocarboxy, thiocarbamoyl, cyano, nitro, nitroso , Azide, hydrazino, ureido, amidino, guanidino, pentafluorothio, trialkylsilyl, alkyl, alkenyl, alkynyl, haloalkyl, alkyloxy, alkenyloxy, alkynyloxy, haloalkyloxy, alkyloxyalkyl, alkyloxyalkyloxy, alkylcarbonyl Alkenylcarbonyl, alkynylcarbonyl, monoalkylamino, dialkylamino, alkylsulfonyl, Kenylsulfonyl, alkynylsulfonyl, monoalkylcarbonylamino, dialkylcarbonylamino, monoalkylsulfonylamino, dialkylsulfonylamino, alkylimino, alkenylimino, alkynylimino, alkylcarbonylimino, alkenylcarbonylimino, alkynylcarbonylimino, alkyloxyimino, Alkenyloxyimino, alkynyloxyimino, alkylcarbonyloxy, alkenylcarbonyloxy, alkynylcarbonyloxy, alkyloxycarbonyl, alkenyloxycarbonyl, alkynyloxycarbonyl, alkylsulfanyl, alkenylsulfanyl, alkynylsulfanyl, alkylsulfinyl, alkenylsulfinyl, and alkynyls Luffy Le.

Further, the “substituted or unsubstituted non-aromatic carbocyclic group” and “substituted or unsubstituted non-aromatic heterocyclic group” may be substituted with “oxo”. In this case, it means a group in which two hydrogen atoms on a carbon atom are substituted as follows.

Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to Formula (IV-viii), Formula (I-i ′) to Formula (I-viii ′), Formula (II-i ′) to Formula (II-viii ′), Formula (III-i ′) to Formula ( III-viii ′), and R ¹ , R ² , R ³ , R ⁴ , R ^5A , R ⁵ , R in the cyclopropane amino acid units represented by formula (IV-i ′) to formula (IV-viii ′) Preferred embodiments of ⁶ , X ^A , X, and Y are shown below.

R ¹ is preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic A carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ¹ is more preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ¹ is particularly preferably hydrogen.
When R ¹ has a substituent, preferred substituents for R ¹ are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ¹ has a substituent, a more preferable substituent of R ¹ is halogen or alkyl.
R ¹ is preferably hydrogen, methyl or ethyl.
R ¹ is more preferably hydrogen.

R ² is preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic A carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ² is more preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ² is particularly preferably hydrogen.
When R ² has a substituent, preferred substituents for R ² are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ² has a substituent, a more preferable substituent of R ² is halogen or alkyl.
R ² is preferably hydrogen, methyl or ethyl.
R ² is more preferably hydrogen.

R ³ is preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic A carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ³ is more preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ³ is particularly preferably hydrogen.
When R ³ has a substituent, preferred substituents for R ³ are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ³ has a substituent, a more preferable substituent of R ³ is halogen or alkyl.
R ³ is preferably hydrogen, methyl or ethyl.
R ³ is more preferably hydrogen.

R ⁴ is preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic A carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ⁴ is more preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ⁴ is particularly preferably hydrogen.
When R ⁴ has a substituent, preferred substituents for R ⁴ are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ⁴ has a substituent, a more preferable substituent of R ⁴ is halogen or alkyl.
R ⁴ is preferably hydrogen, methyl or ethyl.
R ⁴ is more preferably hydrogen.

R ^5A is preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic A carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ^5A is more preferably hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ^5A is particularly preferably hydrogen or substituted or unsubstituted alkyl.
When R ^5A has a substituent, preferred substituents for R ^5A are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ^5A has a substituent, a more preferable substituent of R ^5A is halogen.
R ^5A is preferably hydrogen, ethyl, methyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.
R ^5A is more preferably hydrogen, ethyl, or isobutyl.

R ⁵ is preferably substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic carbocycle A formula group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ⁵ is more preferably substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ⁵ is particularly preferably substituted or unsubstituted alkyl.
When R ⁵ has a substituent, preferred substituents for R ⁵ are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ⁵ has a substituent, a more preferable substituent of R ⁵ is halogen.
R ⁵ is preferably ethyl, methyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.
R ⁵ is more preferably ethyl or isobutyl.

R ⁶ is preferably substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted non-aromatic carbocycle A formula group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group.
R ⁶ is more preferably substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl.
R ⁶ is particularly preferably substituted or unsubstituted alkyl.
When R ⁶ has a substituent, preferred substituents for R ⁶ are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When R ⁶ has a substituent, a more preferable substituent of R ⁶ is halogen.
R ⁶ is preferably ethyl, methyl, n-propyl, isopropyl, n-butyl, isobutyl, or tert-butyl.
R ⁶ is more preferably ethyl.

X ^A is preferably, Fmoc, one protecting group selected Alloc, and the Troc.
X ^A is more preferably Fmoc.

X is preferably hydrogen or one protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc.
X is more preferably one protecting group selected from Boc and Fmoc.
X is particularly preferably Fmoc.

Y is preferably hydrogen, substituted or unsubstituted alkyl, or nucleic acid.
Y is more preferably hydrogen or substituted or unsubstituted alkyl.
Y is particularly preferably hydrogen.
When Y has a substituent, preferred substituents for Y are halogen, cyano, carboxy, carbamoyl, amino, or alkyl.
When Y has a substituent, a more preferable substituent of Y is halogen.
Y is preferably hydrogen, methyl or ethyl.
Y is more preferably hydrogen.

  Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to The compound of formula (IV-viii) is not limited to a particular isomer, but all possible isomers (eg keto-enol isomer, imine-enamine isomer, diastereoisomer, optical Isomers, rotamers etc.), racemates or mixtures thereof.

Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to One or more hydrogen, carbon and / or other atoms of the compound of formula (IV-viii) may be replaced with hydrogen, carbon and / or isotopes of other atoms, respectively. Examples of such isotopes are ² H, ³ H, ¹¹ C, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁸ O, ¹⁷ O, ³¹ P, ³² P, ³⁵ S, ¹⁸ F, ¹²³ I and ^{Like 36} Cl, hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine and chlorine are included. Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to The compound represented by the formula (IV-viii) also includes a compound substituted with such an isotope. The compound substituted with the isotope is also useful as a pharmaceutical, and is represented by formula (Ii) to formula (I-viii), formula (II-i) to formula (II-viii), formula (III-i). ) To formula (III-viii), and all radiolabeled compounds of the compounds represented by formula (IV-i) to formula (IV-viii). In addition, a “radiolabeling method” for producing the “radiolabeled substance” is also encompassed in the present invention, and the “radiolabeled substance” is useful as a metabolic pharmacokinetic study, a research in a binding assay, and / or a diagnostic tool. It is.

Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to The radiolabeled compound of the compound represented by the formula (IV-viii) can be prepared by a method well known in the art. For example, Formula (Ii) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) The tritium-labeled compound represented by the formula (IV-viii) is converted into the formula (Ii) to the formula (I-viii), the formula (II-i) to the formula by the catalytic dehalogenation reaction using tritium. It can be prepared by introducing tritium into a specific compound represented by (II-viii), formula (III-i) to formula (III-viii), and formula (IV-i) to formula (IV-viii). This method is carried out in the presence of a suitable catalyst, for example Pd / C, in the presence or absence of a base, of formula (Ii) to formula (I-viii), formula (II-i) to formula (II). A precursor in which the compounds represented by formula (III-i) to formula (III-viii) and formula (IV-i) to formula (IV-viii) are appropriately substituted with halogen and tritium gas, Including reacting. Other suitable methods for preparing tritium labeled compounds can be referred to “Isotopes in the Physical and Biomedical Sciences, Vol. 1, Labeled Compounds (Part A), Chapter 6 (1987)”. ^{The 14} C-labeled compound can be prepared by using a raw material having ¹⁴ C carbon.

  Formula (I-i) to Formula (I-viii), Formula (II-i) to Formula (II-viii), Formula (III-i) to Formula (III-viii), and Formula (IV-i) to Examples of the pharmaceutically acceptable salt of the compound represented by formula (IV-viii) include, for example, formula (Ii) to formula (I-viii), formula (II-i) to formula (II-viii), A compound represented by formula (III-i) to formula (III-viii) and formula (IV-i) to formula (IV-viii); an alkali metal (for example, lithium, sodium, potassium, etc.), an alkaline earth Metals (eg, calcium, barium, etc.), magnesium, transition metals (eg, zinc, iron, etc.), ammonia, organic bases (eg, trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine) , Triethanolamine, meglumine, diethanolamine, ethylenediamine, pyridine, picoline, quinoline etc.) and salts with amino acids, or inorganic acids (eg hydrochloric acid, sulfuric acid, nitric acid, carbonic acid, hydrobromic acid, phosphoric acid, hydroiodic acid) And organic acids (eg, formic acid, acetic acid, propionic acid, trifluoroacetic acid, citric acid, lactic acid, tartaric acid, oxalic acid, maleic acid, fumaric acid, mandelic acid, glutaric acid, malic acid, benzoic acid, phthalic acid , Ascorbic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, ethanesulfonic acid and the like). Particularly, salts with hydrochloric acid, sulfuric acid, phosphoric acid, tartaric acid, methanesulfonic acid and the like can be mentioned. These salts can be formed by a commonly performed method.

(Method for producing the compound of the present invention)
The compounds represented by formula (I-iii), formula (I-iv), formula (I-vii), and formula (I-viii) according to the present invention are produced, for example, by the general synthesis method shown below. be able to. Extraction, purification, and the like may be performed in a normal organic chemistry experiment.
The compounds of the present invention can be synthesized with reference to techniques known in the art.

Step of Compound a1 → Compound a2 Compound (a2) is obtained by adding (S)-or (R) -2-methyl-2-propanesulfinamide and copper (II) sulfate to the solvent containing compound a1 and reacting. Can do.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 1 hour to 48 hours, preferably 3 hours to 24 hours.
Examples of the reaction solvent include tetrahydrofuran, diethyl ether, dichloromethane, chloroform, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a2 → Compound a3 Compound a3 can be obtained by adding a nucleophile to the solvent containing compound a2 and reacting it. The stereochemistry of the R ⁵ group of the compound a3 obtained depends on the stereochemistry of the sulfinimide in the structural formula of the compound a2 as a raw material. Examples of the nucleophilic agent include lithium reagents such as alkyllithium, Grignard reagents such as alkylmagnesium bromide and alkylmagnesium iodide, and mixed reagents of these and metal salts, and 1 to 10 molar equivalents are used with respect to compound a2. be able to.
The reaction temperature is -78 ° C to the reflux temperature of the solvent, preferably 25 ° C to the reflux temperature.
The reaction time is 30 minutes to 5 hours, preferably 30 minutes to 2 hours.
Examples of the reaction solvent include tetrahydrofuran, hexane, diethyl ether, methyl tert-butyl ether, toluene, dichloromethane and the like, and these can be used alone or in combination.

Step of Compound a3 → Compound a4 Compound a4 can be obtained by adding an acid to a solvent containing compound a3 and reacting.
Examples of the acid include hydrochloric acid-ethyl acetate, hydrochloric acid-dioxane and the like.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include ethyl acetate, dioxane, tetrahydrofuran, diethyl ether, dichloromethane, chloroform, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a4 → Compound a5 Compound a5 can be obtained by reacting a solvent containing compound a4 with a protecting reagent such as a base or an Fmoc-forming reagent. Examples of the base include sodium carbonate, potassium carbonate, sodium hydrogen carbonate and the like. Examples of the Fmoc reagent include 9-fluorenylmethyl N-succinimidyl and 9-fluorenylmethyl chloroformate.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, DMSO, NMP, water, and the like, and these can be used alone or in combination.

Step of Compound a5 → Compound a6 Compound a6 can be obtained by adding an acid to a solvent containing compound a5 and reacting. Examples of the acid include hydrochloric acid-methanol, hydrochloric acid-ethanol, and aqueous hydrochloric acid.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include methanol, ethanol, water and the like, and these can be used alone or in combination.

Step of Compound a6 → Compound a7 Compound a7 can be obtained by adding N-methylmorpholine N-oxide and tetrapropylammonium perruthenate to a solvent containing compound a6 and reacting them.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include dichloromethane, chloroform, tetrahydrofuran, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a7 → Compound a8 Compound a8 can be obtained by adding sulfamic acid and sodium chlorite to a solvent containing compound a7 and reacting them.
The reaction temperature is -20 ° C to 30, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include methanol, ethanol, isopropanol, water and the like, and these can be used alone or in combination.

Compound a8 → step of formula (I-iii), formula (I-iv), formula (I-vii), or formula (I-viii) When Y is a substituted or unsubstituted alkyl, By reacting compound Y—OH in the presence of a condensing agent, formula (I-iii), formula (I-iv), formula (I-vii), or formula (I-viii) can be obtained. .
Examples of the base include DMAP, triethylamine, pyridine, DIEA and the like, and 1 to 5 molar equivalents can be used with respect to compound a8.
As the condensing agent, dicyclohexylcarbodiimide, carbonyldiimidazole, dicyclohexylcarbodiimide-N-hydroxybenzotriazole, EDC, 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methyl Examples thereof include morpholinium chloride and HATU, and 1 to 5 molar equivalents can be used with respect to compound a8.
The reaction temperature is -20 ° C to 60 ° C, preferably 0 ° C to 30 ° C.
The reaction time is 0.1 hour to 24 hours, preferably 1 hour to 12 hours.
Examples of the reaction solvent include DMF, DMA, NMP, tetrahydrofuran, dioxane, dichloromethane, acetonitrile and the like, and these can be used alone or in combination.
When Y is a nucleic acid, Formula (I-iii), Formula (I-iv), Formula (I-vii), or Formula (I-viii) can be obtained by a known technique.

The compounds represented by formula (Ii), formula (I-ii), formula (Iv), and formula (I-vi) according to the present invention are produced, for example, by the general synthesis method shown below. be able to.

Step of compound a4 → compound a9 Compound a9 can be obtained by adding benzyl bromide and potassium carbonate to a solvent containing compound a4 and reacting them.
The reaction temperature is 25 ° C to reflux temperature, preferably 25 ° C to 60 ° C.
The reaction time is 30 minutes to 24 hours, preferably 1 to 5 hours.
Examples of the reaction solvent include DMF, tetrahydrofuran, DMA, DMSO, NMP and the like, and these can be used alone or in combination.

Step of Compound a9 → Compound a10 Compound a10 can be obtained by adding TBAF to a solvent containing compound a9 and reacting.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 1 to 5 hours.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, DMSO, acetonitrile, NMP and the like, and these can be used alone or in combination.

Step of Compound a10 → Compound a11 Compound a11 can be obtained by adding N-methylmorpholine N-oxide and tetrapropylammonium perruthenate to a solvent containing compound a10 and reacting them.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include dichloromethane, chloroform, tetrahydrofuran, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a11 → Compound a12 Compound a12 can be obtained by adding sulfamic acid and sodium chlorite to a solvent containing compound a11 and reacting them.
The reaction temperature is -20 ° C to 30, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include methanol, ethanol, isopropanol, water and the like, and these can be used alone or in combination.

Step of Compound a12 → Compound a13 Compound a13 can be obtained by adding a metal catalyst to a solvent containing compound a12 and reacting with hydrogen gas. Examples of the metal catalyst include palladium-carbon, platinum oxide, rhodium-aluminum oxide, chlorotris (triphenylphosphine) rhodium (I) and the like, and can be used in an amount of 0.01 to 100% by weight based on the compound a12.
As for hydrogen pressure, 1-50 atmospheres is mentioned. As a hydrogen source, cyclohexene, 1,4-cyclohexadiene, formic acid, ammonium formate, or the like can also be used.
The reaction temperature is 0 ° C to the reflux temperature of the solvent, preferably 20 ° C to 40 ° C.
The reaction time is 30 minutes to 120 hours, preferably 1 to 72 hours.
Examples of the reaction solvent include methanol, ethanol, propanol, isopropanol, butanol, tetrahydrofuran, diethyl ether, toluene, ethyl acetate, acetic acid, water and the like, and these can be used alone or in combination.

Step of compound a13 → compound a14 Compound a14 can be obtained by adding a base and a protecting reagent such as an Fmoc reagent to the solvent containing compound a13 for reaction. Examples of the base include sodium carbonate, potassium carbonate, sodium hydrogen carbonate and the like. Examples of the Fmoc reagent include 9-fluorenylmethyl N-succinimidyl and 9-fluorenylmethyl chloroformate.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, DMSO, NMP, water, and the like, and these can be used alone or in combination.

Step of compound a14 → formula (Ii), formula (I-ii), formula (Iv), or formula (I-vi) When Y is substituted or unsubstituted alkyl, By reacting compound Y—OH in the presence of a condensing agent, formula (Ii), formula (I-ii), formula (Iv), or formula (I-vi) can be obtained. .
Examples of the base include DMAP, triethylamine, pyridine, DIEA and the like, and 1 to 5 molar equivalents can be used with respect to compound a14.
As the condensing agent, dicyclohexylcarbodiimide, carbonyldiimidazole, dicyclohexylcarbodiimide-N-hydroxybenzotriazole, EDC, 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methyl Examples thereof include morpholinium chloride and HATU, and 1 to 5 molar equivalents can be used with respect to compound a14.
The reaction temperature is -20 ° C to 60 ° C, preferably 0 ° C to 30 ° C.
The reaction time is 0.1 hour to 24 hours, preferably 1 hour to 12 hours.
Examples of the reaction solvent include DMF, DMA, NMP, tetrahydrofuran, dioxane, dichloromethane, acetonitrile and the like, and these can be used alone or in combination.
When Y is a nucleic acid, Formula (Ii), Formula (I-ii), Formula (Iv), or Formula (I-vi) can be obtained by a known technique.

Formula (II-i), Formula (II-ii), Formula (II-iii), Formula (II-iv), Formula (II-v), Formula (II-vi), Formula (II-) according to the present invention vii), and formula (II-viii), and formula (III-i), formula (III-ii), formula (III-iii), formula (III-iv), formula (III-v), formula (III) The compounds represented by -vi), formula (III-vii), and formula (III-viii) can be produced, for example, by the general synthesis method shown below.

Step of compound a11 → compound a15 Compound a16 can be obtained by adding NaHMDS to a solvent containing (methoxymethyl) triphenylphosphonium chloride and reacting, and further adding compound a12 and conducting a Wittig reaction.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include tetrahydrofuran, dichloromethane, chloroform, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a15 → Compound a16 An aldehyde is obtained by adding an acid to a solvent containing the compound a15. As the acid, hydrochloric acid, sulfuric acid, acetyl chloride and the like can be used.
The reaction temperature is -20 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include tetrahydrofuran, dichloromethane, chloroform, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.
Furthermore, the compound a16 can be obtained by adding sulfamic acid and sodium chlorite to the solvent containing the obtained aldehyde body, and making it react.
The reaction temperature is -20 ° C to 30, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 3 hours.
Examples of the reaction solvent include methanol, ethanol, isopropanol, water and the like, and these can be used alone or in combination.

Step of Compound a16 → Compound a17 A pivaloylated compound can be obtained by adding a base and pivaloyl chloride to a solvent containing the compound a16 for reaction.
Examples of the base include trimethylamine, triethylamine, tripropylamine, tributylamine and the like.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 5 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include dichloromethane, chloroform, tetrahydrofuran, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.
Next, after adding n-butyllithium to a solvent containing (S)-or (R) -4-benzyl-2-oxazolidinone and reacting for 1 to 2 hours, adding a pivaloyl compound and further reacting. Thus, compound a17 can be obtained.
The reaction temperature is -78 ° C to 0 ° C, preferably -78 ° C.
The reaction time is 30 minutes to 3 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include tetrahydrofuran, dichloromethane, chloroform, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a17 → Compound a18 Compound a19 can be obtained by adding NaHMDS to a solvent containing compound a17 and reacting for 1 to 2 hours, and further reacting by adding an alkylating agent. The stereochemistry of the R ⁶ group of the resulting compound a18 depends on the stereochemistry of 4-benzyl-2-oxazolidinone in the structural formula of the compound a17 that is the raw material.
The reaction temperature is -78 ° C to 0 ° C, preferably -78 ° C.
The reaction time is 30 minutes to 3 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include tetrahydrofuran, dichloromethane, chloroform, diethyl ether, benzene, hexane, heptane and the like, and these can be used alone or in combination.

Step of Compound a18 → Compound a19 Compound a19 can be obtained by adding hydrogen peroxide water and lithium hydroxide to a solvent containing compound a18 for reaction.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 1 to 24 hours, preferably 1 to 4 hours.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, acetonitrile, NMP, water and the like, and these can be used alone or in combination.

Step of Compound a19 → Compound a20 Compound a20 can be obtained by adding a metal catalyst to a solvent containing compound a19 and reacting with hydrogen gas.
Examples of the metal catalyst include palladium-carbon, platinum oxide, rhodium-aluminum oxide, chlorotris (triphenylphosphine) rhodium (I), and the like, and can be used in an amount of 0.01 to 100 weight percent with respect to compound a19.
As for hydrogen pressure, 1-50 atmospheres is mentioned. As a hydrogen source, cyclohexene, 1,4-cyclohexadiene, formic acid, ammonium formate, or the like can also be used.
The reaction temperature is 0 ° C to the reflux temperature of the solvent, preferably 20 ° C to 40 ° C.
The reaction time is 30 minutes to 120 hours, preferably 1 to 72 hours.
Examples of the reaction solvent include methanol, ethanol, propanol, isopropanol, butanol, tetrahydrofuran, diethyl ether, toluene, ethyl acetate, acetic acid, water and the like, and these can be used alone or in combination.

Step of Compound a20 → Compound a21 Compound a21 can be obtained by reacting a solvent containing compound a20 with a base and a protecting reagent such as an Fmoc reagent.
Examples of the base include sodium carbonate, potassium carbonate, sodium hydrogen carbonate and the like.
Examples of the Fmoc reagent include 9-fluorenylmethyl N-succinimidyl and 9-fluorenylmethyl chloroformate.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, DMSO, NMP, water, and the like, and these can be used alone or in combination.

Step of Compound a21 → Formula (II-i) to Formula (II-viii), or Formula (III-i) to Formula (III-viii) When Y is substituted or unsubstituted alkyl, By reacting compound Y-OH in the presence of a condensing agent, formula (II-i) to formula (II-viii), or formula (III-i) to formula (III-viii) can be obtained. .
Examples of the base include DMAP, triethylamine, pyridine, DIEA and the like, and 1 to 5 molar equivalents can be used with respect to compound a21.
As the condensing agent, dicyclohexylcarbodiimide, carbonyldiimidazole, dicyclohexylcarbodiimide-N-hydroxybenzotriazole, EDC, 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methyl Examples thereof include morpholinium chloride and HATU, and 1 to 5 molar equivalents can be used with respect to compound a21.
The reaction temperature is -20 ° C to 60 ° C, preferably 0 ° C to 30 ° C.
The reaction time is 0.1 hour to 24 hours, preferably 1 hour to 12 hours.
Examples of the reaction solvent include DMF, DMA, NMP, tetrahydrofuran, dioxane, dichloromethane, acetonitrile and the like, and these can be used alone or in combination.
When Y is a nucleic acid, Formula (II-i) to Formula (II-viii), or Formula (III-i) to Formula (III-viii) can be obtained by a known technique.

Formula (IV-i), Formula (IV-ii), Formula (IV-iii), Formula (IV-iv), Formula (IV-v), Formula (IV-vi), Formula (IV- The compounds represented by vii) and formula (IV-viii) can be produced, for example, by the general synthesis method shown below.

Step of Compound a16 → Compound a22 Compound a22 can be obtained by adding a metal catalyst to a solvent containing compound a16 and reacting with hydrogen gas.
Examples of the metal catalyst include palladium-carbon, platinum oxide, rhodium-aluminum oxide, chlorotris (triphenylphosphine) rhodium (I) and the like, and can be used in an amount of 0.01 to 100% by weight based on the compound a20.
As for hydrogen pressure, 1-50 atmospheres is mentioned. As a hydrogen source, cyclohexene, 1,4-cyclohexadiene, formic acid, ammonium formate, or the like can also be used.
The reaction temperature is 0 ° C to the reflux temperature of the solvent, preferably 20 ° C to 40 ° C.
The reaction time is 30 minutes to 72 hours, preferably 4 to 24 hours.
Examples of the reaction solvent include methanol, ethanol, propanol, isopropanol, butanol, tetrahydrofuran, diethyl ether, toluene, ethyl acetate, acetic acid, water and the like, and these can be used alone or in combination.

Step of Compound a22 → Compound a23 Compound a23 can be obtained by reacting a solvent containing compound a22 with a protecting reagent such as a base or an Fmoc-forming reagent.
Examples of the base include sodium carbonate, potassium carbonate, sodium hydrogen carbonate and the like.
Examples of the Fmoc reagent include 9-fluorenylmethyl N-succinimidyl and 9-fluorenylmethyl chloroformate.
The reaction temperature is 0 ° C to 30 ° C, preferably 0 ° C to 25 ° C.
The reaction time is 30 minutes to 24 hours, preferably 30 minutes to 1 hour.
Examples of the reaction solvent include tetrahydrofuran, DMF, DMA, DMSO, NMP, water, and the like, and these can be used alone or in combination.

Step of Compound a23 → Formula (IV-i) to Formula (IV-viii) When Y is a substituted or unsubstituted alkyl, reacting Compound Y23 with Compound Y—OH in the presence of a base and a condensing agent Thus, formula (IV-i) to formula (IV-viii) can be obtained.
Examples of the base include DMAP, triethylamine, pyridine, DIEA and the like, and 1 to 5 molar equivalents can be used with respect to compound a23.
As the condensing agent, dicyclohexylcarbodiimide, carbonyldiimidazole, dicyclohexylcarbodiimide-N-hydroxybenzotriazole, EDC, 4- (4,6-dimethoxy-1,3,5-triazin-2-yl) -4-methyl Examples thereof include morpholinium chloride and HATU, and 1 to 5 molar equivalents can be used with respect to compound a23.
The reaction temperature is -20 ° C to 60 ° C, preferably 0 ° C to 30 ° C.
The reaction time is 0.1 hour to 24 hours, preferably 1 hour to 12 hours.
Examples of the reaction solvent include DMF, DMA, NMP, tetrahydrofuran, dioxane, dichloromethane, acetonitrile and the like, and these can be used alone or in combination.
When Y is a nucleic acid, Formula (IV-i)-Formula (IV-viii) can be obtained by a well-known method.

The cyclic peptide containing the cyclopropane amino acid unit according to the present invention can be produced, for example, by the following general synthesis method.

(Synthesis of peptide chain)
A chlorotrityl resin (0.5-1.0 mmol / g) fixed with amino acid (AA ¹ ) is suspended in DMF and allowed to swell for 30 minutes, and DMF is removed by filtration. For the amino acid immobilized on the resin, 3 molar equivalents of Fmoc-amino acid (AA ² ), 3 molar equivalents of HBTU, 3 molar equivalents of HOBt, 6 molar equivalents of DIEA are dissolved in DMF. Add the lysate to the resin and shake for 1.5-2 hours. The reaction solution is removed by filtration, and the resin is washed four or more times using DMF. To the amino acid fixed on the resin, 20% piperidine / DMF corresponding to 20 mL / mmol is added and shaken for 20 minutes. The reaction solution is removed by filtration, and the resin is washed four or more times using DMF. The same operation is repeated to synthesize peptide chains AA ^{1 to} AA ⁵ . 1 to 1.5 molar equivalents of Fmoc-cyclopropane amino acid (CP ⁶ ) relative to the amino acid immobilized on the resin, 1 to 2 molar equivalents of HBTU relative to Fmoc-cyclopropane amino acid, and 1 to 2 relative to Fmoc-cyclopropane amino acid Two molar equivalents of DIEA are dissolved in DMF with respect to molar equivalents of HOBt and HBTU. The lysate is added to the resin and shaken for 2-15 hours. The reaction solution is removed by filtration, and the resin is washed four or more times using DMF. To the amino acid fixed on the resin, 20% piperidine / DMF corresponding to 20 ml / mmol is added and shaken for 20 minutes. The reaction solution is removed by filtration, and the resin is washed four or more times with DMF and dichloromethane. For the amino acid immobilized on the resin, 20% hexafluoroisopropanol / dichloromethane corresponding to 30 mL / mmol is added to the resin and shaken for 20 minutes. The reaction is recovered by filtration. The reaction with 20% hexafluoroisopropanol / dichloromethane is repeated twice more, and the reaction solution is recovered by filtration. All of the collected reaction solutions are combined, the solvent is distilled off under reduced pressure, and sufficient drying is performed to obtain a crude product of the protected peptide.

(Cyclization reaction)
DMF is added to the crude product of the protected peptide to a concentration of 0.5 mg / mL and dissolved. For the protected peptide, 1.5 molar equivalents of HOAt, 1.5 molar equivalents of HATU, 1.5 molar equivalents of 2,2,6,6-tetramethylpiperidine, or 4 molar equivalents of DPPA, 6 molar equivalents of Add sodium bicarbonate and stir for 30 minutes to 24 hours. After confirming by LCMS that the starting protected peptide has disappeared, the solvent is distilled off under reduced pressure. The residue is separated using ethyl acetate and saturated aqueous sodium hydrogen carbonate solution, and the solvent of the ethyl acetate layer is distilled off under reduced pressure to obtain a crude product of the cyclic protected peptide.

(Deprotection reaction)
To the crude product of the cyclic protected peptide, add a 10: 1 mixture of TFA and water corresponding to 30 mL / mmol, or a 95: 2.5: 2.5 mixture of TFA, triisopropylsilane, and water. . Stir for 5 hours to react. After confirming the completion of the deprotection reaction by LCMS, the reaction solution is added to an excessive amount of cooled methyl-t-butyl ether, and the resulting precipitate is washed twice or more with the same solvent. In some cases, the solvent of the reaction solution may be distilled off under reduced pressure to obtain a solid. The resulting precipitate is dissolved in DMSO and purified by reverse phase HPLC (0.05% TFA / water-0.05% TFA / acetonitrile or 0.05% TFA / water-0.05% TFA / methanol). By performing, finally, the target cyclic peptide is obtained as a lyophilized powder.

  The present invention will be described in more detail below with reference to Examples, Reference Examples, and Test Examples, but the present invention is not limited thereto.

Moreover, the abbreviation used in this specification represents the following meaning.
Ala: alanine Arg: arginine Asp: aspartic acid Bn: benzyl Cha: cyclohexylalanine DIEA: N, N-diisopropylethylamine DMA: N, N-dimethylacetamide DMF: N, N-dimethylformamide DMSO: dimethyl sulfoxide DPPA: diphenyl phosphate Azide EDC: 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide Leu: leucine Lys: lysine NaHMDS: sodium hexamethyldisilazide NMP: N-methylpyrrolidone TBDPS: tert-butyldiphenylsilyl ether HBTU: O- ( Benzotriazol-1-yl) -N, N, N ′, N′-tetramethyluronium hexafluorophosphate HOBt: 1-hydroxybenzotriazole TBAF Tetrabutylammonium fluoride t-Bu: tert-butyl TFA: trifluoroacetic acid Tyr: Tyrosine

The NMR analysis obtained in each example was performed at 300 MHz and measured using DMSO-d ₆ and CDCl ₃ .

Example 1-1 Synthesis of Compound I-iii-2

Step 1 Synthesis of Compound 2 In a nitrogen atmosphere, Compound 1 (1 g, 2.95 mmol) obtained by the method described in Non-Patent Document 1 or Non-Patent Document 2 was dissolved in toluene (40 mL), and (R)-( +)-2-Methyl-2-propanesulfinamide (716 mg, 5.91 mmol) and copper (II) sulfate (4.71 g, 29.5 mmol) were added, and the mixture was stirred at room temperature for 15 hours. The reaction solution was filtered through Celite, and then the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 2 (1.14 g, yield 87%). .
1H-NMR (500 MHz, CDCl3) δ: 7.65 (d, 6.8 Hz, 4H), 7.56 (d, J = 7.6 Hz, 1H), 7.42 (d, J = 7.1 Hz, 2H), 7.39 (dd, J = 7.1, 6.8Hz, 4H), 3.75 (dd, J = 10.8, 4.9Hz, 1H), 3.64 (dd, J = 11.0, 5.6Hz, 1H), 1.90 (m, 1H), 1.60 (m, 1H), 1.18 (s, 9H), 1.12 (m, 1H), 1.06 (m, 1H), 1.04 (s, 9H).

Step 2 Synthesis of Compound 3 Under a nitrogen atmosphere, compound 2 (958 mg, 2.17 mmol) was dissolved in dichloromethane (22 mL), an ether solution of isobutylmagnesium bromide (1.30 mL, 2.6 mmol) was added, and the mixture was stirred at room temperature for 1 hour. Stir. The reaction mixture was added to saturated ammonium chloride solution and extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 3 (595 mg, yield 55%).
1H-NMR (500 MHz, CDCl3) δ: 7.66 (d, J = 6.5 Hz, 4H), 7.42 (m, 2H), 7.37 (dd, J = 7.1 Hz, 6.5 Hz, 4H), 3.71 (dd, J = 10.6, 5.3 Hz, 1H), 3.48 (dd, J = 10.6, 6.4Hz, 1H), 3.00 (brs, 1H), 2.74 (m, 1H), 1.80 (m, 1H), 1.53 (m, 1H), 1.45 (m, 1H), 1.15 (s, 9H), 1.08 (m, 1H), 1.04 (s, 9H), 0.92 (d, J = 6.4Hz, 3H), 0.90 (d, J = 6.4Hz, 3H ), 0.68 (m, 1H), 0.47 (m, 1H), 0.39 (m, 1H).

Step 3 Synthesis of Compound 4 Under a nitrogen atmosphere, compound 3 (595 mg, 1.19 mmol) was dissolved in ethyl acetate (20 mL), 4 mol / L hydrochloric acid-ethyl acetate solution (9.9 mL, 39.6 mmol) was added, and room temperature was added. For 1 hour. The solvent was distilled off under reduced pressure to obtain a crude product of Compound 4 (499 mg).

Step 4 Synthesis of Compound 5 A crude product of Compound 4 (499 mg) was dissolved in tetrahydrofuran (7.9 mL) and water (4.0 mL), sodium carbonate (505 mg, 4.76 mmol), 9-fluorenylmethyl carbonate. N-succinimidyl (442 mg, 1.31 mmol) was added and stirred at room temperature for 1 hour. Water was added and extracted with ethyl acetate. The extract was washed with saturated brine, dried over anhydrous sodium sulfate, and the solvent was evaporated under reduced pressure to give a crude product of compound 5 (1.16 g).

Step 5 Synthesis of Compound 6 Under a nitrogen atmosphere, a crude product of Compound 5 (1.16 g) was dissolved in methanol (12 mL), and a 4 mol / L hydrochloric acid-ethyl acetate solution (1.79 mL, 7.14 mmol) was added. Stir at room temperature for 1 hour. After the solvent was distilled off under reduced pressure, the obtained residue was purified by silica gel column chromatography (chloroform-ethyl acetate) to obtain Compound 6 (344 mg, yield of 3 steps: 76%).
1H-NMR (500 MHz, CDCl3) δ: 7.77 (d, 7.6 Hz, 2H), 7.59 (d, J = 7.6 Hz, 2H), 7.40 (t, J = 7.6 Hz, 2H), 7.32 (dd, J = 7.6, 7.1Hz, 2H), 4.61 (m, 1H), 4.45 (m, 1H), 4.42 (m, 1H), 4.21 (m, 1H), 3.55 (m, 1H), 3.27 (m, 1H), 3.14 (m, 1H), 1.67 (m, 1H), 1.41 (m, 2H), 1.16 (m, 1H), 0.94-0.86 (m, 6H), 0.73 (m, 1H), 0.48 (m, 2H) .

Step 6 Synthesis of Compound 7 Compound 6 (342 mg, 0.90 mmol) was dissolved in dichloromethane (6.8 mL) under a nitrogen atmosphere, and molecular sieves (171 mg), N-methylmorpholine N-oxide (211 mg, 1.8 mmol) were dissolved. , Tetrapropylammonium perruthenate (15.8 mg, 0.045 mmol) was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was filtered through celite, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform-ethyl acetate) to obtain Compound 7 (308 mg, yield 91%).
1H-NMR (500MHz, CDCl3) δ: 9.24 (brs, 1H), 7.77 (d, J = 7.0Hz, 2H), 7.57 (d = 7.0Hz, 2H), 7.40 (t, J = 7.0Hz, 2H) , 7.32 (t, J = 7.0Hz, 2H), 4.48 (m, 1H), 4.45 (m, 1H), 4.21 (m, 1H), 3.38 (m, 1H), 2.01 (m, 1H), 1.65 ( m, 1H), 1.53 (m, 1H), 1.42 (m, 1H), 1.40 (m, 1H), 1.31 (m, 1H), 1.00 (m, 1H), 0.94-0.85 (m, 6H).

Step 7 Synthesis of Compound I-iii-2 Compound 7 (306 mg, 0.81 mmol) was dissolved in methanol (12.2 mL), water (3.1 mL), sulfamic acid (118 mg, 1.22 mmol), chlorous acid Sodium (110 mg, 1.22 mmol) was added and stirred at 0 ° C. for 1 hour. 2 mol / L hydrochloric acid was added and extracted with chloroform. The extract was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The obtained residue was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound I-iii-2 (231 mg, yield 72%).
1H-NMR (500 MHz, CDCl3) δ: 7.76 (d, J = 7.6 Hz, 2H), 7.58 (d, d = 7.0, 2H), 7.40 (t, J = 7.6 Hz, 2H), 7.31 (t, J = 7.0Hz, 2H), 4.54 (m, 1H), 4.43 (m, 1H), 4.21 (m, 1H), 3.34 (m, 1H), 1.73 (m, 1H), 1.65 (m, 1H), 1.50 (m, 1H), 1.41 (m, 1H), 1.26 (m, 1H), 0.94-0.85 (m, 6H), 0.74 (m, 1H), 0.53 (m, 1H).

The following compound I-iii-1, compound I-iv-1, and compound I-iv-2 were also synthesized by the same synthesis method.

Example 1-2 Synthesis of Compound I-i-2

Step 1 Synthesis of Compound 10 In a nitrogen atmosphere, Compound 9 (5.0 g, 14.8 mmol) obtained by the method described in Non-Patent Documents 1 and 2 was dissolved in toluene (200 mL), and (S)-(+) 2-Methyl-2-propanesulfinamide (3.58 g, 29.5 mmol) and copper (II) sulfate (23.6 g, 148 mmol) were added, and the mixture was stirred at room temperature for 15 hours. The reaction mixture was filtered through celite, and the solvent was evaporated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to give compound 10 (6.45 g, yield 99%). .
1H-NMR (600MHz, CDCl3) δ: 7.76 (d, J = 8.3Hz, 1H), 7.67-7.63 (m, 4H), 7.43-7.35 (m, 6H), 3.95 (dd, J = 11.3, 6.2 Hz , 1H), 3.69 (dd, J = 11.3, 8.0Hz, 1H), 2.10 (m, 1H), 1.69 (m, 1H), 1.24 (m, 1H), 1.18 (s, 9H), 1.03-0.98 ( m, 10H).

Step 2 Synthesis of Compound 11 Under a nitrogen atmosphere, Compound 17 (6.45 g, 14.6 mmol) was dissolved in toluene (1.16 L), and a tetrahydrofuran solution of isobutylmagnesium bromide (29.2 mL, 58.4 mmol) was added. Stir at 110 ° C. for 30 minutes. The reaction mixture was added to saturated ammonium chloride solution and extracted with ethyl acetate. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 11 (3.74 g, yield 51%).
1H-NMR (600 MHz, CDCl3) δ: 7.71-7.65 (m, 4H), 7.42 (m, 2H), 7.38 (m, 4H), 3.84 (dd, J = 11.1, 6.4 Hz, 1H), 3.72 (dd, J = 11.1, 7.0Hz, 1H), 2.86 (m, 1H), 1.81 (m, 2H), 1.47 (m, 1H), 1.19 (m, 1H), 1.08 (s, 9H), 1.07 ( s, 9H), 1.06 (m, 1H), 0.88 (d, J = 6.4Hz, 3H), 0.87 (d, J = 6.4Hz, 3H), 0.73 (m, 1H), 0.10 (m, 1H).

Step 3 Synthesis of Compound 12 Under a nitrogen atmosphere, Compound 11 (3.74 g, 7.48 mmol) was dissolved in ethyl acetate (125 mL), a 4 mol / L hydrochloric acid-ethyl acetate solution (61.7 mL, 247 mmol) was added, and room temperature was added. For 1 hour. After the solvent was distilled off under reduced pressure, the resulting residue was purified by amino silica gel column chromatography (chloroform-methanol) to obtain a crude product of compound 12 (3.38 g, 97%).

Step 4 Synthesis of Compound 13 Under a nitrogen atmosphere, a crude product of Compound 12 (3.38 g, 7.48 mmol) was dissolved in DMF (89 mL), benzyl bromide (2.67 mL, 22.4 mmol), potassium carbonate (3 0.62 g, 26.2 mmol) was added, and the mixture was stirred at 60 ° C. for 1 hour. The reaction solution was added to water, extracted with ethyl acetate, and the extract was dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 13 (3.83 g, yield 89%).
1H-NMR (600MHz, CDCl3) δ: 7.66 (dd, J = 7.6, 1.2Hz, 2H), 7.61 (dd, J = 8.2, 1.7Hz, 2H), 7.45-7.37 (m, 4H), 7.33 (t , J = 7.6, 2H), 7.18-7.15 (m, 4H), 7.12-7.09 (m, 6H), 4.30 (dd, J = 11.1, 4.7Hz, 1H), 3.64 (d, J = 13.5, 2H) , 3.39 (d, J = 13.5, 2H), 2.93 (dd, J = 11.1, 10.0Hz, 1H), 1.99 (m, 1H), 1.78 (m, 1H), 1.28 (m, 1H), 1.21 (m , 1H), 1.06 (s, 9H), 0.88 (m, 1H), 0.84 (m, 1H), 0.74 (d, J = 6.4Hz, 3H), 0.68 (d, J = 7.0Hz, 3H), 0.66 (m, 1H), -0.31 (m, 1H).

Step 5 Synthesis of Compound 14 Under a nitrogen atmosphere, compound 13 (3.83 g, 6.65 mmol) was dissolved in tetrahydrofuran (57.5 mL), a TBAF solution in tetrahydrofuran (16.6 mL, 16.6 mmol) was added, and at room temperature. Stir for 3 hours. The reaction mixture was added to saturated brine, extracted with ethyl acetate, and the extract was dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 14 (1.92 g, yield 86%).
1H-NMR (600MHz, CDCl3) δ: 7.36 (d, J = 7.0Hz, 4H), 7.30 (dd, J = 7.6, 7.0Hz, 4H), 7.24 (t, J = 7.6Hz, 2H), 5.81 ( d, J = 11.1Hz, 1H), 3.98 (d, J = 12.3Hz, 2H), 3.83 (m, 1H), 3.32 (d, J = 12.3Hz, 2H), 2.58 (t, J = 11.4Hz, 1H), 2.34 (m, 1H), 1.85 (m, 1H), 1.68 (m, 1H), 1.52 (m, 1H), 1.32 (m, 1H), 1.00 (d, J = 6.4Hz, 3H), 0.95 (m, 1H), 0.82 (m, 1H), 0.69 (d, J = 6.4Hz, 3H), 0.05 (m, 1H).

Step 6 Synthesis of Compound 15 Under a nitrogen atmosphere, Compound 14 (1.92 g, 5.69 mmol) was dissolved in dichloromethane (38.4 mL), molecular sieves (960 mg), N-methylmorpholine N-oxide (1.33 g, 11.4 mmol) and tetrapropylammonium perruthenate (100 mg, 0.28 mmol) were added and stirred at room temperature for 1 hour. The reaction mixture was filtered through celite, and the solvent was evaporated under reduced pressure. The obtained residue was purified by silica gel column chromatography (chloroform-ethyl acetate) to obtain a crude product of compound 15 (1.87 g).

Step 7 Synthesis of Compound 16 A crude product of Compound 15 (58 mg, 0.17 mmol) was dissolved in methanol (2.3 mL) and water (0.58 mL), sulfamic acid (25.2 mg, 0.26 mmol), Sodium chlorate (23.5 mg, 0.26 mmol) was added and stirred at 0 ° C. for 1 hour. Saturated aqueous ammonium chloride solution was added, the mixture was extracted with chloroform, and the extract was washed with saturated brine. The extract was dried over anhydrous sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 16 (33 mg, 55%).
1H-NMR (600 MHz, CDCl3) δ: 7.38 (d, J = 7.0 Hz, m, 4H), 7.32 (dd, J = 7.3, 7.0 Hz, 4H), 7.28 (m, 2H), 4.09 (d, J = 12.9Hz, 2H), 3.61 (d, J = 12.9Hz, 2H), 2.84 (m, 1H), 1.83 (m, 1H), 1.78 (m, 1H), 1.72 (m, 1H), 1.62 (m , 1H), 1.23 (m, 1H), 1.00 (d, J = 7.0Hz, 3H), 0.94 (m, 1H), 0.80 (m, 1H), 0.73 (d, J = 6.4Hz, 3H).

Step 8 Synthesis of Compound 17 Compound 16 (33 mg, 0.094 mmol) was dissolved in methanol (1.3 mL), hydrous palladium hydroxide (11 mg, 0.0078 mmol) was added, and the mixture was stirred under a hydrogen gas atmosphere for 6 hours. . The reaction mixture was filtered through celite, and the solvent was evaporated under reduced pressure to give a crude product of compound 17 (17 mg).

Step 9 Synthesis of Compound I-i-2 Crude product of Compound 17 (17 mg, 0.094 mmol) was dissolved in tetrahydrofuran (0.62 mL) and water (0.31 mL), and sodium carbonate (30 mg, 0.28 mmol). 9-Fluorenylmethyl N-succinimidyl carbonate (31.7 mg, 0.094 mmol) was added and stirred at 0 ° C. for 2 hours. 2 mol / L hydrochloric acid was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (chloroform-methanol) to obtain Compound Ii-2 (36 mg, yield of 97% for two steps).
1H-NMR (500MHz, DMSO-d6) δ: 7.89 (d, 7.3Hz, 2H), 7.68 (dd, J = 7.5, 7.3Hz, 2H), 7.41 (t, J = 7.5Hz, 2H), 7.33 ( m, 2H), 7.20 (d, J = 8.5Hz, 1H), 4.25 (m, 1H), 4.19 (m, 2H), 3.53 (m, 1H), 1.59 (m, 2H), 1.50 (m, 1H ), 1.39 (m, 1H), 1.30 (m, 1H), 1.03 (m, 1H), 0.88-0.84 (m, 6H), 0.81 (m, 1H).

The following compound I-i-1 and compound I-ii-1 were also synthesized by the same synthesis method.

Example 1-3 Synthesis of Compound II-i-1

Step 1 Synthesis of Compound 19 Under a nitrogen atmosphere, (methoxymethyl) triphenylphosphonium chloride (4.21 g, 12.3 mmol) was dissolved in tetrahydrofuran (33.0 mL) and cooled to 0 ° C. A 1.1 mol / L-NaHMDS tetrahydrofuran solution (13.4 mL, 14.8 mmol) was added and stirred at 0 ° C. for 1 hour, and then a tetrahydrofuran solution (0.15 mL, 4.4) of the compound 15 obtained in Example 2 was used. 9 mmol) was added and the mixture was further stirred at 0 ° C. for 1 hour. Saturated aqueous ammonium chloride solution was added, the mixture was extracted with ethyl acetate, and the extract was washed with saturated brine. The extract was dried over anhydrous sodium sulfate, and the solvent was distilled off under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain a crude product of compound 19 (1.38 g). It was.

Step 2 Synthesis of Compound 20 A crude product (1.38 g) of Compound 19 was dissolved in tetrahydrofuran (34.5 mL) and water (34.5 mL), acetyl chloride (8.13 mL, 114 mmol) was added, and 0 Stir for 1 hour at ° C. The reaction solution was added to saturated aqueous sodium hydrogen carbonate solution and extracted with chloroform, and the extract was washed with saturated brine. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain a residue.
The obtained residue was dissolved in methanol (55.2 mL) and water (13.8 mL), sulfamic acid (553 mg, 5.69 mmol) and sodium chlorite (515 mg, 5.69 mmol) were added, and Stir for hours. Saturated aqueous ammonium chloride solution was added, the mixture was extracted with chloroform, and the extract was washed with saturated brine. After the extract was dried over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure, and the resulting residue was purified by silica gel column chromatography (chloroform-methanol) to obtain compound 20 (1.07 g, 77%).
1H-NMR (600 MHz, CDCl3) δ: 7.50-7.41 (m, 4H), 7.37-7.31 (m, 6H), 4.12 (m, 2H), 3.53 (m, 2H), 2.56 (dd, J = 14.1, 1.8Hz, 1H), 2.52 (dd, J = 14.1, 2.3Hz, 1H), 1.91 (m, 1H), 1.78 (m, 1H), 1.72 (m, 1H), 1.15 (m, 1H), 1.06 ( d, J = 7.0Hz, 3H), 1.05 (m, 1H), 1.00 (m, 1H), 0.91 (m, 1H), 0.77 (d, J = 6.4Hz, 3H), -0.08 (m, 1H) .

Step 3 Synthesis of Compound 21 Compound 20 (360 mg, 0.99 mmol) was dissolved in dichloromethane (9.9 mL) under a nitrogen atmosphere, and then pivaloyl chloride (0.16 mL, 1.28 mmol), triethylamine (0.18 mL, 1 .28 mmol) was added and the mixture was stirred at 0 ° C. for 1 hour. Water was added and the mixture was extracted with dichloromethane, and the extract was washed with saturated brine. After drying over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain a residue of pivaloylated product.
Under a nitrogen atmosphere, (S) -4-benzyl-2-oxazolidinone (558 mg, 3.15 mmol) was dissolved in tetrahydrofuran (9.9 mL) and cooled to -78 ° C. A 2.6 mol / L-n-butyllithium hexane solution (1.14 mL, 2.95 mmol) was added, and the mixture was stirred at -78 ° C for 1 hour. To this was added the tetrahydrofuran solution of the pivaloylated compound obtained earlier, and the mixture was further stirred at -78 ° C for 1 hour. Acetic acid was added to the reaction mixture, diluted with saturated brine, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 21 (300 mg, 58%).
1H-NMR (500 MHz, CDCl3) δ: 7.37-7.34 (m, 4H), 7.33 (m, 2H), 7.30-7.24 (m, 5H), 7.22 (m, 2H), 7.19 (m, 2H), 4.68 (m, 1H), 4.2 (t, J = 9.0Hz, 1H), 4.17 (dd, J = 9.0, 3.7Hz, 1H), 3.78 (d, J = 13.7Hz, 2H), 3.54 (d, J = 13.7Hz, 2H), 3.41 (dd, J = 16.5, 4.8Hz, 1H), 3.31 (dd, J = 13.3, 3.3Hz, 1H), 2.78 (dd, J = 12.3, 9.3Hz, 1H), 2.37- 2.30 (m, 2H), 1.90 (m, 1H), 1.68 (m, 1H), 1.36 (m, 1H), 1.17 (m, 1H), 0.96 (m, 1H), 0.85 (d, J = 6.6Hz , 3H), 0.84-0.80 (m, 5H).

Step 4 Synthesis of Compound 22 Compound 21 (297 mg, 0.57 mmol) was dissolved in tetrahydrofuran (5.9 mL) under a nitrogen atmosphere and cooled to −78 ° C. 1.1 mol / L-NaHMDS tetrahydrofuran solution (0.67 mL, 0.74 mmol) was added and stirred at −78 ° C. for 1 hour, then methyl iodide (0.35 mL, 5.7 mmol) was added, and further −78 ° C. For 1 hour. Acetic acid was added to the reaction solution, diluted with water, extracted with chloroform, and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (ethyl acetate-hexane) to obtain Compound 22 (247 mg, 81%).
1H-NMR (600MHz, CDCl3) δ: 7.36 (t, J = 7.0Hz, 2H), 7.31-7.28 (m, 5H), 7.24-7.19 (m, 6H), 7.12 (t, J = 8.2Hz, 2H ), 4.45 (m, 1H), 3.97 (dd, J = 9.4, 2.3Hz, 1H), 3.70 (d, J = 14.1Hz, 2H), 3.61 (d, J = 7.6Hz, 1H), 3.58 (d , J = 14.1Hz, 2H), 3.23 (dd, J = 13.5, 3.5Hz, 1H), 3.14 (m, 1H), 2.71 (dd, J = 13.5, 10.0Hz, 1H), 2.51 (q, J = 7.0Hz, 1H), 1.94 (m, 1H), 1.46 (m, 1H), 1.40 (m, 1H), 1.37 (d, J = 7.0Hz, 3H), 1.1 (m, 1H), 1.01 (m, 1H), 0.79 (d, J = 7.6Hz, 3H), 0.78 (m, 1H), 0.63 (d, J = 6.4Hz, 3H), 0.06 (m, 1H).

Step 5 Synthesis of Compound 23 Compound 22 (247 mg, 0.46 mmol) was dissolved in tetrahydrofuran (2.5 mL) and water (0.5 mL), and 30% aqueous hydrogen peroxide (0.23 mL, 2.29 mmol), 1 mol / L lithium hydroxide (0.92 mL, 0.92 mmol) was added, and the mixture was stirred at 0 ° C. for 15 hours. The reaction solution was added to a saturated aqueous sodium sulfate solution and extracted with chloroform. The extract was washed successively with 2 mol / L hydrochloric acid and saturated brine, and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by silica gel column chromatography (chloroform-methanol) to obtain Compound 23 (164 mg, 94%).
1H-NMR (600 MHz, CDCl3) δ: 7.65-7.50 (m, 2H), 7.43-7.22 (m, 8H), 4.28 (m, 1H), 3.87 (m, 1H), 3.59 (m, 1H), 3.43 (m, 1H), 2.49 (td, J = 10.2, 1.8Hz, 1H), 1.935 (m, 1H), 1.77 (m, 1H), 1.72 (m, 1H), 1.06 (d, J = 6.4Hz, 3H), 1.02 (d, J = 4.8Hz, 3H), 1.02-0.94 (m, 3H), 0.91 (m, 1H), 0.82 (d, J = 6.4Hz, 3H), -0.12 (m, 1H) .

Step 6 Synthesis of Compound 24 Compound 23 (149 mg, 0.39 mmol) was dissolved in methanol (6.0 mL), hydrous palladium hydroxide (46 mg, 0.033 mmol) was added, and the mixture was stirred under a hydrogen gas atmosphere for 4 days. . The reaction mixture was filtered through celite, and the solvent was evaporated under reduced pressure to obtain a crude product of compound 58 (87 mg).

Step 7 Synthesis of Compound II-i-1 A crude product of Compound 24 (87 mg, 0.39 mmol) was dissolved in tetrahydrofuran (2.4 mL) and water (1.2 mL), and sodium carbonate (125 mg, 1.18 mmol) was dissolved. 9-fluorenylmethyl N-succinimidyl carbonate (146 mg, 0.43 mmol) was added and stirred at 0 ° C. for 4 hours. 2 mol / L hydrochloric acid was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by carboxyl group-modified silica gel column chromatography (ethyl acetate-hexane) to obtain Compound II-i-1 (136 mg, 2-step yield 82%).
1H-NMR (600 MHz, DMSO-d6) δ: 7.88 (d, J = 7.6 Hz, m, 2H), 7.71 (d, J = 7.6 Hz, 1H), 7.65 (d, J = 7.0 Hz, 1H), 7.41 (t, J = 7.3Hz, 2H), 7.33 (d, J = 7.3, 1H), 7.31 (d, J = 7.3Hz, 1H), 7.06 (d, J = 9.4Hz, 1H), 4.22-4.17 (m, 2H), 4.16 (m, 1H), 3.06 (m, 1H), 2.04 (m, 1H), 1.55 (m, 1H), 1.45 (m, 1H), 1.32 (m, 1H), 1.17 ( d, J = 7.0Hz, 3H), 1.03-0.94 (m, 2H), 0.84 (d, J = 6.7Hz, 3H), 0.83 (d, J = 6.7Hz, 3H) ,, 0.68 (m, 1H) , -0.07 (m, 1H).

The following compound III-ii-1, compound II-iii-1, compound III-iv-1, compound II-i-2, III-ii-2 and II-iii-2 were synthesized in the same manner.

Example 1-4 Synthesis of Compound IV-i-1

Step 1 Synthesis of Compound 26 Compound 20 (100 mg, 0.27 mmol) was dissolved in methanol (4.0 mL), hydrous palladium hydroxide (50 mg, 0.036 mmol) was added, and the mixture was stirred under a hydrogen gas atmosphere for 15 hours. . The reaction solution was filtered through celite, and then the solvent was distilled off under reduced pressure to obtain a crude product (54 mg) of Compound 26.

Step 2 Synthesis of Compound IV-i-1 A crude product of Compound 26 (51 mg, 0.27 mmol) was dissolved in tetrahydrofuran (1.52 mL) and water (0.76 mL), and sodium carbonate (87 mg, 0.82 mmol). 9-fluorenylmethyl N-succinimidyl carbonate (92 mg, 0.27 mmol) was added and stirred at 0 ° C. for 3 hours. 2 mol / L hydrochloric acid was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by distilling off the solvent under reduced pressure was purified by carboxyl group-modified silica gel column chromatography (ethyl acetate-hexane) to obtain Compound IV-i-1 (88 mg, 2-step yield 79%).
1H-NMR (600 MHz, DMSO-d6) δ: 7.89 (d, J = 7.6 Hz, m, 2H), 7.69 (t, J = 7.0 Hz, 2H), 7.41 (t, J = 7.6 Hz, 2H), 7.32 (m, 2H), 7.20 (d = 9.0, 1H), 4.31 (m, 2H), 4.20 (m, 1H), 2.98 (m, 1H), 2.47 (m, 1H), 2.39 (dd, J = 16.5, 5.3Hz, 1H), 1.56 (m, 1H), 1.47 (m, 1H), 1.30 (m, 1H), 0.99 (m, 1H), 0.89 (m, 1H), 0.85 (d, J = 6.6 Hz, 3H), 0.83 (d, J = 6.6Hz, 3H), 0.72 (m, 1H), -0.01 (m, 1H).

The following compound IV-iii-1 was also synthesized in the same manner.

Example 1-5 Synthesis of Compound I-1

Step 1 Synthesis of Compound 29 Compound 28 (230 mg, 2.0 mmol) obtained by the method described in Non-Patent Document 3 was dissolved in tetrahydrofuran (4.6 mL) and water (2.3 mL), and sodium carbonate (636 mg, 6 0.0 mmol), 9-fluorenylmethyl carbonate N-succinimidyl (810 mg, 2.4 mmol) was added and stirred at room temperature for 1 hour. 2 mol / L hydrochloric acid was added, and the mixture was extracted with chloroform. The extract was washed with saturated brine and dried over anhydrous sodium sulfate. The residue obtained by evaporating the solvent under reduced pressure was purified by carboxyl group-modified silica gel column chromatography (ethyl acetate-hexane) to obtain Compound I-1 (630 mg, yield 93%).
1H-NMR (500 MHz, CD3OD) δ: 7.79 (d, 7.6 Hz, 2H), 7.64 (d, J = 7.3 Hz, 2H), 7.38 (t, J = 7.3 Hz, 2H), 7.31 (t, J = 7.6Hz, 2H), 4.35 (d, J = 6.8Hz, 2H), 4.21 (t, J = 6.8Hz, 1H), 3.15 (dd, J = 14.1, 6.0Hz, 1H), 3.02 (dd, J = 14.1, 6.8Hz, 1H), 1.57 (m, 1H), 1.51 (m, 1H), 1.10 (m, 1H), 0.86 (m, 1H).

The following compound I-2 was also synthesized by the same method.

Example 2-1 Peptide No. 5 synthesis (synthesis of peptide chain)
A chlorotrityl resin (0.60 mmol / g, 0.028 mmol, 47 mg) immobilized with Arg was suspended in DMF and allowed to swell for 30 minutes, and DMF was removed by filtration. Dissolve Fmoc-Asp (t-Bu) -OH (35 mg, 0.084 mmol), HBTU (32 mg, 0.084 mmol), HOBt-H ₂ O (13 mg, 0.084 mmol), DIEA (29 μL, 0.168 mmol) The DMF solution (0.21 mL) was added to the resin and shaken for 2 hours. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (0.56 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed four times with DMF. The same operation was sequentially performed using Fmoc-Cha-OH and Fmoc-Lys (Boc) -OH to synthesize a peptide chain Lys-Cha-Asp-Arg.
Compounds were dissolved in DMF of 0.45mL I-iii-1 (20mg , 0.056mmol), HBTU (42mg, 0.112mmol), HOBt-H 2 O (17mg, 0.112mmol), DIEA (40μL, 0. 224 mmol) was added to the resin synthesized peptide chain and shaken for 2 hours. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (0.56 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed 4 times with DMF and 4 times with dichloromethane.
After adding 0.8% TFA / dichloromethane (0.56 mL) to the resin and shaking for 3 minutes, the reaction solution was collected by filtration. The same operation was further repeated 4 times, and the reaction solution was recovered by filtration. After all the collected reaction solutions were combined, they were neutralized using DIEA (0.11 mL), and the solvent was distilled off under reduced pressure. The residue was sufficiently dried to obtain a crude product (31 mg) of the protected peptide.

(Cyclization reaction)
31 mg of the obtained crude product of the protected peptide was dissolved in 6.2 mL of DMF, HOAt (11.6 mg, 0.085 mmol) dissolved in 0.06 mL of DMF, and HATU dissolved in 0.06 mL of DMF (32 .4 mg, 0.085 mmol) and DIEA (30 μL, 0.17 mmol) were sequentially added. After stirring the reaction solution for 1 hour, the solvent was distilled off under reduced pressure. The obtained residue was separated using a 10% acetonitrile aqueous solution and dichloromethane, and the dichloromethane layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain 41 mg of a crude product of cyclic protected peptide.

(Deprotection reaction)
41 mg of the resulting crude product of the cyclic protected peptide was dissolved in a 95: 2.5: 2.5 mixed solution (0.56 mL) of TFA, triisopropylsilane, and water, and stirred for 2 hours. The reaction solution was added to cooled isopropyl ether (12 mL), and the resulting precipitate was washed twice with isopropyl ether. The precipitate was dissolved in a small amount of DMSO, purified by reverse phase HPLC (0.05% TFA / water-0.05% TFA / acetonitrile), and peptide No. 5 was obtained as a lyophilized powder. 5 (11.6 mg, 61.5% yield) was obtained.

Example 2-2 Peptide No. Synthesis of 12 (synthesis of peptide chain)
A chlorotrityl resin (0.82 mmol / g, 0.09 mmol, 110 mg) fixed with Leu was suspended in DMF and allowed to swell for 30 minutes, and DMF was removed by filtration. Fmoc-Leu-OH (95 mg, 0.27 mmol), HBTU (102 mg, 0.27 mmol), HOBt-H ₂ O (41 mg, 0.27 mmol), DIEA (94 μL, 0.54 mmol) in DMF (0.45 mL) ) Was added to the resin and shaken for 1 hour. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (2 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed four times with DMF.
The same operation was sequentially performed using Fmoc-Ala-OH, Fmoc-Leu-OH, and Fmoc-Tyr (t-Bu) -OH to synthesize a peptide chain Tyr-Leu-Ala-Leu-Leu.
Compounds were dissolved in DMF of 0.45mL II-i-1 (38mg , 0.09mmol), HBTU (68mg, 0.18mmol), HOBt-H 2 O (27mg, 0.18mmol), DIEA (63μL, 0. 36 mmol) was added to the resin synthesized peptide chain and shaken for 2 hours. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (2 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed 4 times with DMF and 4 times with dichloromethane.
After adding 20% hexafluoroisopropanol / dichloromethane (3 mL) to the resin and shaking for 20 minutes, the reaction solution was collected by filtration. The reaction was further performed twice using 20% hexafluoroisopropanol / dichloromethane, and the reaction solution was recovered by filtration. All the collected reaction solutions were combined, the solvent was distilled off under reduced pressure, and the residue was sufficiently dried to obtain a crude product (75 mg) of the protected peptide.

(Cyclization reaction)
The obtained crude purified product of protected peptide (75 mg) was dissolved in 150 mL of DMF, HOAt (18.4 mg, 0.14 mmol) dissolved in 0.5 mL of DMF, and HATU (51.3 mg) dissolved in 0.5 mL of DMF. 0.14 mmol) and 2,2,6,6-tetramethylpiperidine (23 μL, 0.14 mmol) were added sequentially. After stirring the reaction solution for 30 minutes, the solvent was distilled off under reduced pressure. The obtained residue was separated using a 10% acetonitrile aqueous solution and dichloromethane, and the dichloromethane layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain 69 mg of a crude product of cyclic protected peptide.

(Deprotection reaction)
69 mg of the resulting crude product of the cyclic protected peptide was dissolved in a 10: 1 mixture of TFA and water (3 mL) and stirred for 30 minutes. After the solvent of the reaction solution was distilled off under reduced pressure, the residue was dissolved in a small amount of DMSO and purified by reverse phase HPLC (0.05% TFA / water-0.05% TFA / acetonitrile). 12 (8.4 mg, 12.4% yield) was obtained.

Example 2-3 Peptide No. Synthesis of 23 (synthesis of peptide chain)
A chlorotrityl resin (0.94 mmol / g, 0.047 mmol, 50 mg) fixed with Ala was suspended in DMF and allowed to swell for 30 minutes, and DMF was removed by filtration. Fmoc-Ala-OH (44mg, 0.141mmol), HBTU (54mg, 0.141mmol), HOBt-H 2 O (22mg, 0.141mmol), DIEA (49μL, 0.282mmol) DMF solution (0 obtained by dissolving .24 mL) was added to the resin and shaken for 1 hour. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (0.94 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed four times with DMF.
The same operation was sequentially performed using Fmoc-Leu-OH, Fmoc-Ala-OH, and Fmoc-Tyr (t-Bu) -OH to synthesize a peptide chain Tyr-Ala-Leu-Ala-Ala.
Compound II-i-1 (20 mg, 0.047 mmol), HBTU (18 mg, 0.047 mmol), HOBt-H ₂ O (7.2 mg, 0.047 mmol), DIEA (16 μL, dissolved in 0.24 mL of DMF 0.094 mmol) was added to the resin synthesized peptide chain and shaken for 2 hours. The reaction solution was removed by filtration, and the resin was washed four times with DMF. 20% piperidine / DMF (0.94 mL) was added to the resin and shaken for 20 minutes. The reaction solution was removed by filtration, and the resin was washed 4 times with DMF and 4 times with dichloromethane.
After adding 20% hexafluoroisopropanol / dichloromethane (1.4 mL) to the resin and shaking for 20 minutes, the reaction solution was collected by filtration. The reaction was further performed twice using 20% hexafluoroisopropanol / dichloromethane, and the reaction solution was recovered by filtration. All the collected reaction solutions were combined, the solvent was distilled off under reduced pressure, and the residue was sufficiently dried to obtain a crude product (36 mg) of the protected peptide.

(Cyclization reaction)
36 mg of the crude product of the obtained protected peptide was dissolved in 70 mL of DMF, HOAt (12.8 mg, 0.094 mmol) dissolved in 0.7 mL of DMF, and HATU (35.7 mg) dissolved in 0.7 mL of DMF. 0.094 mmol) and 2,2,6,6-tetramethylpiperidine (16 μL, 0.094 mmol) were sequentially added. After stirring the reaction solution for 30 minutes, the solvent was distilled off under reduced pressure. The obtained residue was separated using a 10% acetonitrile aqueous solution and dichloromethane, and the dichloromethane layer was dried over anhydrous sodium sulfate. The solvent was distilled off under reduced pressure to obtain 37 mg of a crude product of cyclic protected peptide.

(Deprotection reaction)
37 mg of the resulting crude product of the cyclic protected peptide was dissolved in a 10: 1 mixture of TFA and water (0.75 mL) and stirred for 30 minutes. After the solvent of the reaction solution was distilled off under reduced pressure, the residue was dissolved in a small amount of DMSO and purified by reverse phase HPLC (0.05% TFA / water-0.05% TFA / acetonitrile). 23 (4.2 mg, yield 13.4%) was obtained.

The cyclic peptides and the cyclic peptides into which cyclopropane amino acid units were introduced are summarized in the following table. In addition, the cyclic peptide in the table | surface which is not described in the said Example was synthesize | combined by the method similar to the cyclic peptide of the said Example. In the table, “/” in the peptide sequence indicates that a cyclic peptide is formed by amide bonding at both ends (N-terminal and C-terminal) of the peptide.

Test Example 1 Metabolic stability test (serum stability test)
To a mixed solution of bovine serum (45 μL) and 10 mmol / L phosphate buffer (pH 7.4, 45 μL), an aqueous solution (10 μL) in which the cyclic peptides 1 to 6 are dissolved at a concentration of 5 mg / mL, respectively, is added. It was allowed to stand for a certain time at ° C. The reaction solution (10 μL) was diluted with an 80% acetonitrile aqueous solution (50 μL) containing 0.05% TFA, and the supernatant was analyzed by HPLC after centrifugation. The residual ratio of the peptide was measured from the peak area of HPLC.

(Trypsin stability test)
An aqueous solution in which the cyclic peptides 1 to 6 are dissolved at a concentration of 5 mg / mL in a 10 mmol / L phosphate buffer solution (pH 7.4, 90 μL) in which bovine pancreas-derived trypsin is dissolved at a concentration of 0.5 mg / mL ( 10 μL) was added and allowed to stand at 25 ° C. for a fixed time. The reaction solution (10 μL) was diluted with an 80% acetonitrile aqueous solution (50 μL) containing 0.05% TFA, and the supernatant was analyzed by HPLC after centrifugation. The residual ratio of the peptide was measured from the peak area of HPLC.

(Protease stability test)
An aqueous solution (10 μL) in which the peptides 1 to 6 are dissolved at a concentration of 5 mg / mL in 10 mmol / L phosphate buffer (pH 7.4, 90 μL) in which a protease derived from Streptomyces grieseus is dissolved at a concentration of 0.5 mg / mL. ) Was added and allowed to stand at 37 ° C. for a fixed time. The reaction solution (10 μL) was diluted with an 80% acetonitrile aqueous solution (50 μL) containing 0.05% TFA, and the supernatant was analyzed by HPLC after centrifugation. The residual ratio of the peptide was measured from the peak area of HPLC.

The measurement results are shown below. The numerical value represents the remaining rate (%). Peptide No. 3, 4, and 5 are peptide Nos. Into which no cyclopropane amino acid unit was introduced. Compared with 2, the metabolic stability was particularly improved.

Test Example 2 HIV Integrase LEDGF Inhibitory Activity Test Whether a cyclopropane amino acid unit was introduced into a cyclic peptide (peptide No. 2) having HIV integrase LEDGF inhibitory activity was tested.

  Various concentrations of 100% DMSO-dissolved peptide solution were dispensed at 0.2 μL / well into a 384-well black plate. A solution prepared in 20.4 nM 6 × His-HIV-integrase and 10.2 nM Anti-6 × His-d2 was dispensed at 9.8 μL / well. The peptide solution and the HIV-integrase solution were mixed and allowed to stand at room temperature (23 ° C.) for 20 minutes. A solution prepared in 6 nM Flag-LEDGF and 1 nM Anti-FLAG-EuK was dispensed at 10 μL / well. A total of 20 μL / well of the reaction solution was reacted at room temperature (23 ° C.) for 2 hours or more. By performing HTRF signal detection (Excitation: 320 nm, Emission: 665, 615 nm) with an HTRF reagent (an anti-FLAG antibody converted to europium cryptee travel and an anti-6xHis antibody labeled with d2), the cyclic peptides 1 to 6 are used. The binding ability inhibition ability was measured.

The results are shown below. In particular, peptide no. Compared with the cyclic peptide (peptide No. 2) before introduction, activity was maintained in the wild type strain and increased in the mutant strain.

Test Example 3 Membrane Permeability Test The LLC-PK1 cells used in the membrane permeability test were prepared using medium A [Medium 199 (manufactured by Invitrogen), FBS (10%), Gentamycin (0.05 mg / mL), hyromycin C (100 μg / mL). mL)] at 37 ° C. in the presence of 5% carbon dioxide / 95% oxygen. These cells were implanted in Transwell insert (96 holes, pore size 0.4 μm) at a cell density of 1.4 × 10 4 cells / insert, and medium B (Medium 199 (manufactured by Invitrogen), FBS (10%), Gentamycin ( 0.05 mg / mL)] was added. These cells were further cultured for 4-6 days at 37 ° C. in the presence of 5% carbon dioxide / 95% oxygen and used for the membrane permeability test.
The culture solution on the apical membrane side was replaced with a buffer solution containing each of the peptides 7 to 23 at a concentration of 20 μmol / L, and then cultured at 37 ° C. for 1 hour. The buffer solutions on the apical membrane side and the basement membrane side were collected, the drug concentration in each buffer solution was quantified by LC / MS / MS, and the Pe value was calculated.

Pe = permeation drug amount (pmol) / cell membrane area (cm ² ) / initial concentration (nM) / culture time (sec)

The results are shown below. The higher the Pe value, the higher the membrane permeability. As a reference example, cLogP and Pe values of cyclosporine, which is a cyclic peptide having high membrane permeability, are also shown. From this result, it was found that peptide No. in which no cyclopropane amino acid unit was introduced. Compared to 7, a plurality of peptides introduced with cyclopropane amino acid units showed improved membrane permeability.

  Since the membrane permeability and / or metabolic stability of the cyclic peptide can be improved by introducing the cyclopropane amino acid unit according to the present invention into the cyclic peptide, the cyclic peptide containing the cyclopropane amino acid unit according to the present invention is Useful as a medicine.

Claims (8)

The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbon A cyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group)
A method for improving the membrane permeability and / or metabolic stability of the cyclic peptide by selecting any of the cyclopropane amino acid units represented by the above formula and introducing the cyclopropane amino acid unit into the ring of the cyclic peptide.
The following formula:

(The symbols in the formula are as defined in claim 1)
Is introduced into the ring of the cyclic peptide, and the cyclic peptide before introduction of the cyclopropane amino acid unit is compared with membrane permeability and / or metabolic stability. For selecting a cyclopropane amino acid unit that improves the membrane permeability and / or metabolic stability of.
The following formula:

(Where
R ¹ , R ² , R ³ , and R ⁴ are each independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, or substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic A carbocyclic group, a substituted or unsubstituted non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ^5A is independently hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted unsubstituted An aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
R ⁵ and R ⁶ are each independently substituted or unsubstituted alkyl, substituted or unsubstituted alkenyl, substituted or unsubstituted alkynyl, substituted or unsubstituted aromatic carbocyclic group, substituted or unsubstituted A non-aromatic carbocyclic group, a substituted or unsubstituted aromatic heterocyclic group, or a substituted or unsubstituted non-aromatic heterocyclic group,
X ^A is one protecting group selected from Fmoc, Alloc, and Troc;
X is hydrogen or one protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc;
Y is hydrogen, substituted or unsubstituted alkyl, or nucleic acid)
Any of the compounds shown below (excluding the following compounds)

).
Any compound of formula (II-i)-(II-viii), (III-i)-(III-viii), and (IV-i)-(IV-viii), wherein X is 4. A compound according to claim 3 which is hydrogen.
Any compound of formula (II-i)-(II-viii), (III-i)-(III-viii), and (IV-i)-(IV-viii), wherein X is 4. The compound of claim 3, which is a protecting group selected from Boc, Cbz, Fmoc, Alloc, and Troc.
The compound according to any one of claims 3 to 5, wherein Y is hydrogen.
The following formula:

(The symbols in the formula are as defined in claim 1)
The cyclic peptide which has either group shown by in a ring.
The following formula:

The cyclic peptide which has either group shown by in a ring.