阅读说明：本技术 用于制备环状有机化合物的方法 (Process for preparing cyclic organic compounds ) 是由 本间晶江 侯增烨 伊藤久师 筱仓洁 于 2018-05-11 设计创作，主要内容包括：本发明的目的在于提供使用连续搅拌釜式反应器(CSTR)制备环状有机化合物的方法,所述方法能够实现优异的杂质抑制效果(质量改善)、反应釜尺寸的减小、连续生产等。为了解决上述问题,本发明人对使用CSTR的环化反应进行了研究,该反应器并未常规地用于环状化合物的环化反应。因此,本发明人已经发现与常规环化方法相比,本发明可以实现优异的杂质抑制效果(质量改善)、反应釜尺寸的减小、连续生产等。此外,本发明人已经发现,通过将常规上主要以精细化学品工厂水平使用的模拟方法应用于本发明的环化反应,由此实验性地预测环化反应的反应速率,并且在使用CSTR的环化反应中设定影响这些条件的流量(停留时间)、前体和环状有机化合物的浓度以及用于环化反应的温度等,甚至在环肽和杂环化合物的制备中可以有效地实现上述改善效果。(An object of the present invention is to provide a method for preparing a cyclic organic compound using a Continuous Stirred Tank Reactor (CSTR), which is capable of achieving excellent impurity suppression effect (quality improvement), reduction in reactor size, continuous production, and the like. In order to solve the above problems, the present inventors have studied on a cyclization reaction using a CSTR, which is not conventionally used for a cyclization reaction of a cyclic compound. Therefore, the present inventors have found that the present invention can achieve an excellent impurity suppressing effect (quality improvement), reduction in the size of a reaction vessel, continuous production, and the like, as compared with the conventional cyclization method. Furthermore, the present inventors have found that the above-described improvement effect can be effectively achieved even in the production of cyclic peptides and heterocyclic compounds by applying a simulation method conventionally used mainly at the fine chemical plant level to the cyclization reaction of the present invention, thereby experimentally predicting the reaction rate of the cyclization reaction, and setting the flow rate (residence time), the concentrations of the precursor and cyclic organic compound, and the temperature for the cyclization reaction, etc., which affect these conditions, in the cyclization reaction using CSTR.)

1. A process for the preparation of a cyclic organic compound, said process comprising a cyclisation reaction step of cyclisation of a cyclisation precursor of said cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR).

2. The method of claim 1, wherein the cyclic organic compound is a peptide compound comprising a cyclic moiety, wherein the compound is comprised of natural amino acids and/or amino acid analogs.

3. The method according to claim 1 or 2, wherein the peptidal compound comprising a cyclic moiety comprises a cyclic moiety consisting of 4 to 14 natural amino acid and/or amino acid analogue residues, and wherein the total number of natural amino acid and amino acid analogue residues is 7 to 20.

4. The method of claim 3, wherein the cyclic organic compound has the following characteristics:

(i) comprising at least two N-substituted amino acids and at least one non-N-substituted amino acid; and

(ii) has a ClogP value of 6 or more.

5. The method of any one of claims 1 to 4, wherein the cyclization reaction is via an intramolecular cyclization reaction of one or more bonds selected from the group consisting of:

(i) an amide bond;

(ii) a disulfide bond;

(iii) an ether bond;

(iv) a thioether bond;

(v) an ester linkage;

(vi) a thioester bond; and

(vii) a carbon-carbon bond.

6. The method according to claim 1 or 2, wherein the cyclic organic compound is a compound represented by the following general formula (I):

(wherein X represents CH or N; R1 represents a hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C1-4 haloalkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group, a C1-6 alkoxy group or a C1-6 alkylthio group; R2 represents a hydrogen atom, a halogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group, or R2 and R3 together form a ring; R3 represents a hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group or a C1-6 alkoxy group, or R2 and R3 together form a ring; R4 represents a hydrogen atom, a halogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group; R5, R6 and R39 7 each independently represents a hydrogen atom, a halogenOptionally substituted C1-6 alkylene, C2-6 alkenylene, C2-6 alkynylene, C3-10 cycloalkylene, C3-10 cycloalkenylene, C6-12 arylene, or-3 to 12 membered monocyclic heterocycle-; l1, L2 and L3 each independently represent a single bond, -CONR8-, -NR8CO-, -NR8-, -O-, -SO₂NR8-、-NR8SO₂-, -COO-, NR8CONR 8' -, NR8 COO-or-OCONR 8-; and R8 and R8' each independently represent a hydrogen atom or an optionally substituted C1-6 alkyl group).

7. The process according to any one of claims 1 to 6, wherein the cyclization reaction is carried out on an industrial scale using conditions obtained based on the results of preliminary tests of the cyclization reaction.

8. The method of claim 7, wherein the condition is obtained by a step comprising:

(i) in the preliminary test, concentration change data with time at a plurality of temperatures is obtained for at least one selected from the group consisting of: the cyclization precursor, the cyclic organic compound, one or more intermediates and one or more by-products;

(ii) (ii) determining a reaction rate constant k by using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n；

(iii) (iii) by using the temperature used in step (i), the reaction rate constant k determined in step (ii)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iv) By using the frequency factor A determined in step (iii)_nAnd activation energy E_nEquation (II) above and the reaction rate equation above, determine the reaction rate constant k at the cyclization temperature in one or more CSTRs_n(ii) a And

(v) (iii) use of the reaction rate constant k determined in step (iv)_nThe above reaction rate equation and the following CSTR mass balance equation (III) to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

9. The method of claim 8, wherein the promoiety of the cyclization reaction is represented by the following formula (IV):

and a reaction rate constant k₁、k₂And k₃Determined using any one of the following equations (V) to (IX):

(wherein TM represents a cyclic organic compound, SM represents a cyclized precursor, ACT represents an activator, IM represents an intermediate, Dimer represents a Dimer, and C represents a concentration (M)).

10. The method of claim 8, wherein the promoiety of the cyclization reaction is represented by the following formula (X):

and a reaction rate constant k₁And k2 is determined using any of the following equations (XI) to (XIII):

(where r denotes reaction rate, TM denotes target molecule, SM denotes cyclized precursor (═ intermediate), ACT denotes activator, Dimer denotes Dimer, and C denotes concentration (M)).

11. The method of any one of claims 7 to 10, wherein the condition is selected from the group consisting of: a flow rate in the continuous stirred tank reactor, a concentration of the cyclization precursor, and a concentration of the cyclic organic compound.

12. A process for promoting the intramolecular cyclization of a cyclization precursor, said process comprising the step of cyclizing a cyclization precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR).

13. A process for obtaining conditions for the cyclisation precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) on an industrial scale, comprising the following steps:

(i) in the preliminary test, concentration change data over time at a plurality of temperatures is obtained for at least one selected from the group consisting of: the cyclization precursor, the cyclic organic compound, one or more intermediates and one or more by-products;

(ii) (ii) determining a reaction rate constant k using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n；

(iii) (iii) by using the temperature used in step (i), the reaction rate constant k determined in step (ii)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iv) By using the frequency factor A determined in step (iii)_nAnd activation energy E_nThe above equation (II) and the above reaction rate equation, the reaction rate constant k at the cyclization temperature in the CSTR is determined_n(ii) a And

(v) (iii) use of the reaction rate constant k determined in step (iv)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

14. A program for causing a computer to carry out the following steps (i) to (iv) to obtain conditions for the cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR):

(i) the reaction rate constant k was determined by using_n：

Data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: said cyclization precursor, said cyclic organic compound, one or more intermediates and one or more by-products, and

a reaction rate equation associated with the cyclization reaction;

(ii) (ii) by using the temperature used in step (i), the reaction rate constant k determined in step (i)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iii) By using the frequency factor A determined in step (ii)_nAnd activation energy E_nThe above equation (II) and the above reaction rate equation, the reaction rate constant k at the cyclization temperature in the CSTR is determined_n(ii) a And

(iv) (iv) use of the reaction rate constant k determined in step (iii)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

15. A system for obtaining conditions for the cyclisation precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) in an industrial scale cyclisation, the system comprising:

(i) the reaction rate constant k was determined by using_nThe device of (2):

data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: said cyclization precursor, said cyclic organic compound, one or more intermediates and one or more by-products, and

a reaction rate equation associated with the cyclization reaction;

(ii) (ii) by using the temperature used in step (i), the reaction rate constant k determined in step (i)_nAnd the following equation (II) to determine the frequency factor A_nAnd activation energy E_nThe device of (2):

(iii) by using the frequency factor A determined in step (ii)_nAnd activation energy E_nThe reaction rate constant k at the cyclization temperature in the CSTR was determined by the above equation (II) and the above reaction rate equation_nThe apparatus of (1); and

(iv) (iv) use of the reaction rate constant k determined in step (iii)_nThe above reaction rate equation and the following CSTR mass balance equation (III) to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

Technical Field

The present invention relates to a process for the synthesis of macrocyclic organic compounds using a continuous stirred tank reactor.

Background

The macrocyclic compound (cyclic compound) refers to a compound including a heterocyclic compound (non-patent document 1 and patent document 1) and a peptide compound (patent document 2). Macrocyclic compounds are used as natural/non-natural bioactive compounds for use in pharmaceuticals and the like. In order to provide such drugs, the preparation of cyclic compounds is an important task for the pharmaceutical industry. The preparation of the cyclic compound cannot be completed without a cyclization step. Cyclization of a compound refers to a reaction in which a single molecule reacts at more than two reactive sites within the molecule to form a cyclic structure. The combination mode is different; for example, cyclization reactions through various bonds such as an amide bond, an ester bond, an ether bond, a thioether bond, and a disulfide bond are known (non-patent documents 2 and 3, and patent documents 3 and 4).

It is known that in the cyclization reaction, an increase in concentration causes competition between intramolecular reactions and intermolecular reactions, resulting in the production of dimers or higher-order polymeric compounds due to the intermolecular reactions. Although the concentration can be kept relatively high in usual polymer synthesis, the reaction for cyclizing a single molecule is performed under dilution conditions to suppress the production of a polymer caused by an intermolecular reaction (non-patent document 4 and patent documents 1, 5, 6, and 7). This limits the maximum throughput per batch depending on the size of the reactor. Meanwhile, methods for simulating dilution conditions are known, such as a method of adding a reaction substrate dropwise to a solution in small portions under reaction conditions (reverse dropwise addition), and a method of using a solid-phase-supported substrate in a reaction (on-resin cyclization) (non-patent document 5 and patent documents 8 and 9). These methods allow obtaining the cyclic compounds of interest while avoiding dilution conditions; however, these methods still have limitations in throughput depending on the size of the reaction tank and the nature of the reaction substrate.

A Continuous Stirred Tank Reactor (CSTR) (or mixed flow reactor) refers to a tank reactor that operates continuously. CSTRs are widely used for liquid phase reactions, gas phase reactions, and heterogeneous reactions, like batch reactors in which tank reactors are operated in a batch manner (non-patent document 6). Ideally, the reaction liquid in the tank reactor is sufficiently mixed by stirring so that the temperature and concentration in the tank become uniform, and the reaction liquid is allowed to flow out of the tank at the same concentration and temperature as the inside of the reactor (mixed flow type). Further, the tubular reactor is a reaction apparatus equipped with a single tube or a plurality of tubes placed in parallel, and produces a concentration distribution in the direction of the tube axis (plug flow type). Both CSTRs and tubular reactors are continuous reactors suitable for continuous production. CSTRs are common reactors in the field of fine chemicals, and are known for polymerization control of polymer molecules (patent document 10), enzymatic reactions (patent document 11), and the like. Further, it is also known to combine a CSTR with other generally known continuous operation techniques such as continuous liquid-liquid separation and crystallization, utilizing the continuous operability of the CSTR (non-patent document 7).

Summary of The Invention

Problems to be solved by the invention

Continuous reactors for large-scale production in the fine chemicals field have never been used for the cyclization of cyclic organic compounds (including heterocyclic compounds) in the pharmaceutical field, in particular of cyclic peptide drugs. There may be a number of reasons behind this; for example, many commercially available cyclic peptide drugs have high activity and thus are not in high demand for large-scale production.

Continuous Stirred Tank Reactors (CSTRs) and tubular reactors are known continuous reactors. In a tubular reactor, the substrate and product concentrations at the inlet and outlet are different due to the principle of the reactor. In the case of a tubular reactor, when a heterocyclic compound, a peptide compound, or the like is cyclized, a relatively dilute solution needs to flow through the reactor, and thus a large amount of an organic solvent is required. In contrast, in the case of CSTR, the distribution of the concentrations of the cyclization precursor and the cyclization product can be controlled to be uniform in the reactor, and by adjusting the residence time or the like, the concentration of the cyclization precursor in the reactor can be kept low while the amount of the solvent used is reduced. The present inventors have paid attention to this point.

The present invention has been accomplished in view of the above circumstances. An object of the present invention is to provide a process for producing a cyclic organic compound using a CSTR, which can achieve an excellent impurity-suppressing effect (quality improvement, for example, suppression of intermolecular cyclization reaction), size reduction of one or more reaction vessels, continuous production, and the like, as compared with conventional cyclization processes.

Means for solving the problems

In order to solve the above problems, the present inventors studied a cyclization reaction using a Continuous Stirred Tank Reactor (CSTR) which is not conventionally used for a cyclization reaction of a cyclic compound, thereby finding that such a method can achieve an excellent impurity suppression effect (quality improvement), size reduction of a reaction tank, continuous production, and the like, as compared with a conventional cyclization method, and thus completed the present invention.

Furthermore, the present inventors found that the above-described improvement effect can be effectively achieved in the production of cyclic peptides or heterocyclic compounds by applying a simulation method conventionally used mainly at the fine chemical plant level to the cyclization reaction of the present invention, thereby experimentally predicting the reaction rate of the cyclization reaction, and setting the flow rate (residence time), the concentrations of the precursor and the cyclic organic compound, and the temperature for the cyclization reaction and the like that affect these conditions in the cyclization reaction using CSTR.

The present invention is based on such a finding, and specifically provides the following [1] to [15 ]:

[1] a process for the preparation of a cyclic organic compound, said process comprising a cyclisation reaction step of cyclisation of a cyclisation precursor of said cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR);

[2] the method according to [1], wherein the cyclic organic compound is a peptide compound comprising a cyclic moiety, wherein the compound is composed of natural amino acids and/or amino acid analogues;

[3] the method according to [1] or [2], wherein the peptide compound comprising a cyclic moiety comprises a cyclic moiety consisting of 4 to 14 natural amino acid and/or amino acid analogue residues, and wherein the total number of natural amino acid and amino acid analogue residues is 7 to 20;

[4] the method according to [3], wherein the cyclic organic compound has the following characteristics:

(i) comprising at least two N-substituted amino acids and at least one non-N-substituted amino acid; and

(ii) has a ClogP value of 6 or greater;

[5] the method according to any one of [1] to [4], wherein the cyclization reaction is an intramolecular cyclization reaction through one or more bonds selected from the group consisting of:

(i) an amide bond;

(ii) a disulfide bond;

(iii) an ether bond;

(iv) a thioether bond;

(v) an ester linkage;

(vi) a thioester bond; and

(vii) a carbon-carbon bond;

[6] the method according to [1] or [2], wherein the cyclic organic compound is a compound represented by the following general formula (I):

(wherein X represents CH or N; R1 representsA hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C1-4 haloalkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group, a C1-6 alkoxy group or a C1-6 alkylthio group; r2 represents a hydrogen atom, a halogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group, or R2 and R3 together form a ring; r3 represents a hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group or a C1-6 alkoxy group, or R2 and R3 together form a ring; r4 represents a hydrogen atom, a halogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group; r5, R6 and R7 each independently represent optionally substituted C1-6 alkylene, C2-6 alkenylene, C2-6 alkynylene, C3-10 cycloalkylene, C3-10 cycloalkenylene, C6-12 arylene or 3-to 12-membered monocyclic heterocycle-; l1, L2 and L3 each independently represent a single bond, -CONR8-, -NR8CO-, -NR8-, -O-, -SO₂NR8-、-NR8SO₂-, -COO-, NR8CONR 8' -, NR8 COO-or-OCONR 8-; and R8 and R8' each independently represent a hydrogen atom or an optionally substituted C1-6 alkyl group);

[7] the process according to any one of [1] to [6], wherein the cyclization reaction is carried out on an industrial scale using conditions obtained based on the results of preliminary tests of the cyclization reaction;

[8] the method according to [7], wherein the condition is obtained by a step comprising:

(i) in the preliminary test, concentration change data with time at a plurality of temperatures is obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) determining a reaction rate constant k by using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n；

(iii) (iii) by using the temperature used in step (i), the reaction rate constant k determined in step (ii)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iv) By using the frequency factor A determined in step (iii)_nAnd activation energy E_nThe above equation (II) and the above reaction rate equation, the reaction rate constant k at the cyclization temperature in CSTR is determined_n(ii) a And

(v) (iii) use of the reaction rate constant k determined in step (iv)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration);

[9] the method according to [8], wherein the motif reaction of the cyclization reaction is represented by the following formula (IV):

and a reaction rate constant k₁、k₂And k₃Determined using any one of the following equations (V) to (IX):

(wherein TM represents a cyclic organic compound, SM represents a cyclized precursor, ACT represents an activator, IM represents an intermediate, Dimer represents a Dimer, and C represents a concentration (M));

[10] the method according to [8], wherein the motif reaction of the cyclization reaction is represented by the following formula (X):

and a reaction rate constant k₁And k₂Determined using any of the following equations (XI) through (XIII):

(where r represents the reaction rate, TM represents the target molecule, SM represents the cyclized precursor (═ intermediate), ACT represents the activator, Dimer represents the Dimer, and C represents the concentration (M));

[11] the method according to any one of [7] to [10], wherein the condition is selected from the group consisting of: the flow rate in the continuous stirred tank reactor, the concentration of the cyclization precursor, and the concentration of the cyclic organic compound;

[12] a process for promoting the intramolecular cyclization of a cyclization precursor, said process comprising the step of cyclizing a cyclization precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR);

[13] a process for obtaining conditions for the cyclisation precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) on an industrial scale, comprising the following steps:

(i) in the preliminary test, concentration change data with time at a plurality of temperatures was obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) determining a reaction rate constant k using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n；

(iii) (iii) by using the temperature used in step (i), the reaction rate constant k determined in step (ii)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iv) By using the frequency factor A determined in step (iii)_nAnd activation energy E_nThe above equation (II) and the above reaction rate equation, the reaction rate constant k at the cyclization temperature in the CSTR is determined_n(ii) a And

(v) (iii) use of the reaction rate constant k determined in step (iv)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration);

[14] a program for causing a computer to carry out the following steps (i) to (iv) to obtain conditions for the cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR):

(i) determination of the reaction Rate constant k_nBy using:

data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates and one or more by-products, and

a reaction rate equation associated with the cyclization reaction;

(ii) (ii) by using the temperature used in step (i), the reaction rate constant k determined in step (i)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(iii) By using the frequency factor A determined in step (ii)_nAnd activation energy E_nThe above equation (II) and the above reaction rate equation, the reaction rate constant k at the cyclization temperature in the CSTR is determined_n(ii) a And

(iv) (iv) use of the reaction rate constant k determined in step (iii)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration); and

[15] a system for obtaining conditions for the cyclisation precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) in an industrial scale cyclisation, the system comprising:

(i) the reaction rate constant k was determined by using the following concentration variation data and reaction rate equation relating to the cyclization reaction_nThe device of (2):

data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) by using the temperature used in step (i), the reaction rate constant k determined in step (i)_nAnd the following equation (II) to determine the frequency factor A_nAnd activation energy E_nThe device of (2):

(iii) by using the frequency factor A determined in step (ii)_nAnd activation energy E_nThe reaction rate constant k at the cyclization temperature in the CSTR was determined by the above equation (II) and the above reaction rate equation_nThe apparatus of (1); and

(iv) (iv) use of the reaction rate constant k determined in step (iii)_nThe above reaction rate equation and the following CSTR mass balance equation (III) to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

In addition, the following invention is also provided:

[2-1] A method for suppressing an intermolecular reaction of a cyclized precursor, which comprises the step of cyclizing a cyclized precursor of a cyclic organic compound in at least one continuous stirred tank reactor;

[2-2] use of a continuous stirred tank reactor for intramolecular cyclization of a cyclization precursor of a cyclic organic compound in at least one continuous stirred tank reactor;

[2-3] the method according to [7], wherein the condition is obtained by a step comprising:

(i) in the preliminary test, concentration change data with time at a single temperature was obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) determining a reaction rate constant k by using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n(ii) a And

(iii) (iii) use of the reaction rate constant k determined in step (ii)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration);

[2-4] A process for obtaining conditions for the cyclisation of a cyclic precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) on an industrial scale, comprising the following steps:

(i) in the preliminary test, concentration change data with time at a single temperature was obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) determining a reaction rate constant k by using the concentration variation data obtained in step (i) and a reaction rate equation relating to the cyclization reaction_n(ii) a And

(iii) (iii) use of the reaction rate constant k determined in step (ii)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration);

[2-5] A program for causing a computer to carry out the following steps to obtain conditions for the cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR):

(i) the reaction rate constant k was determined by using the following concentration variation data and reaction rate equation relating to the cyclization reaction_n：

Data obtained in preliminary tests on the change in concentration over time at a single temperature for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products; and

(ii) (ii) Using the reaction Rate constant k determined in step (i)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Represents a supply concentration, and C represents a concentration); and

[2-6] A system for obtaining conditions for the cyclisation precursor of a cyclic organic compound in at least one continuous stirred tank reactor on an industrial scale, said system comprising:

(i) the reaction rate constant k was determined by using the following concentration variation data and reaction rate equation relating to the cyclization reaction_nThe device of (2):

obtained in preliminary tests, for at least one selected from the group consisting of: concentration data for a cyclization precursor, a cyclic organic compound, one or more intermediates, and one or more side products over time at a single temperature; and

(ii) (ii) Using the reaction Rate constant k determined in step (i)_nThe above reaction rate equation and the following CSTR mass balance equation (III), means to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

Effects of the invention

The present invention provides a process for producing a cyclic organic compound by performing a cyclization reaction using one or more CSTRs, which can achieve an excellent impurity suppression effect, a reduction in the size of a reaction vessel, continuous production, and the like, as compared with conventional cyclization processes. Further, the present invention provides a production method that can more effectively achieve the above-described improvement effect by applying a simulation method to determine conditions for the cyclization reaction.

Brief Description of Drawings

FIG. 1 shows a hypothetical apparatus for the synthesis of 2kg of target molecule using a CSTR (conditions of run 4 in example 1).

Fig. 2 shows a hypothetical apparatus for the synthesis of 2kg of target molecule by pseudo high dilution (also called high dilution) (conditions of run 3 in example 1).

Figure 3 shows the 1H NMR pattern of a sample of cyclosporin a cyclised precursor (example 2).

FIG. 4 shows the equipment for making cyclosporin A using a CSTR in example 3.

FIG. 5 shows the 1H NMR chart of a sample of desmopressin (example 4).

Figure 6 shows the 1H NMR chart of a sample of the cyclized precursor of desmopressin (example 4).

Fig. 7 shows the 1H NMR pattern of a sample of cyclosporin a derivative (example 5).

Figure 8 shows the 1H NMR pattern of a sample of the cyclised precursor of cyclosporin a derivative (example 5).

Modes for carrying out the invention

< definition of substituents etc. >

As used herein, the term "alkyl" refers to a monovalent group derived by removing any one hydrogen atom from an aliphatic hydrocarbon and having a subset of hydrocarbon groups or hydrocarbon group structures that contain neither heteroatoms nor unsaturated carbon-carbon bonds in the backbone, but rather hydrogen and carbon atoms. The carbon chain length n is in the range of 1 to 20, and the alkyl group is preferably a C2-C10 alkyl group. Examples of the alkyl group include "C1-C6 alkyl" and specifically include methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, tert-butyl, sec-butyl, 1-methylpropyl, 1-dimethylpropyl, 2, 2-dimethylpropyl, 1, 2-trimethylpropyl, 1, 2, 2-trimethylpropyl, 1, 2, 2-tetramethylpropyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2, 2-dimethylbutyl, 2, 3-dimethylbutyl, 3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, isopentyl, and neopentyl.

Herein, the term "alkenyl" refers to a monovalent group having at least one double bond (two adjacent SP2 carbon atoms). Depending on the arrangement of the double bonds and substituents, if they are present, the double bonds may take a trans-lateral (entgegen) (E) or an ipsilateral (zusammen) (Z) and cis-or trans-geometric form. Examples of alkenyl groups include straight or branched chains, including straight chains containing internal olefins. Preferred examples thereof include C2-C10 alkenyl groups, and more preferably C2-C6 alkenyl groups.

Specific examples of alkenyl groups include vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl (including cis-and trans-forms), 3-butenyl, pentenyl, and hexenyl.

Herein, the term "alkynyl" refers to a monovalent group having at least one triple bond (two adjacent SP carbon atoms). Examples thereof include straight-chain or branched-chain alkynyl groups (including internal alkylene groups). Preferred examples thereof include C2-C10 alkynyl groups, and more preferably C2-C6 alkynyl groups.

Specific examples of the alkynyl group include ethynyl, 1-propynyl, propargyl, 3-butynyl, pentynyl, hexynyl, 3-phenyl-2-propynyl, 3- (2' -fluorophenyl) -2-propynyl, 2-hydroxy-2-propynyl, 3- (3-fluorophenyl) -2-propynyl and 3-methyl- (5-phenyl) -4-pentynyl.

As used herein, the term "cycloalkyl" means a saturated or partially saturated, cyclic, monovalent aliphatic hydrocarbon group containing a monocyclic, bicyclic, or spiro ring. Preferred examples thereof include C3-C10 cycloalkyl groups. Specific examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and bicyclo [2.2.1] heptyl.

Herein, the term "aryl" means a monovalent aromatic hydrocarbon ring. Preferred examples thereof include C6-C10 aryl groups. Specific examples of the aryl group include phenyl and naphthyl groups (e.g., 1-naphthyl and 2-naphthyl).

In this context, the term "heteroaryl" means an aromatic cyclic monovalent group which preferably contains from 1 to 5 heteroatoms in the ring-forming atoms and which may be partially saturated. The rings may be monocyclic or bicyclic fused rings (e.g., bicyclic heteroaryl groups formed by fusion with benzene or monocyclic heteroaryl groups). The number of ring-forming atoms is preferably 5 to 10 (5-to 10-membered heteroaryl).

Specific examples of heteroaryl groups include furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiadiazolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, benzofuranyl, benzothienyl, benzothiadiazolyl, benzothiazolyl, benzoxazolyl, benzooxadiazolyl, benzimidazolyl, indolyl, isoindolyl, indazolyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, quinoxalinyl, benzodioxolyl, indolizinyl, and imidazopyridinyl groups.

Herein, the term "arylalkyl (aralkyl)" means a group containing both aryl and alkyl groups, for example, a group derived from the above alkyl groups by substituting at least one hydrogen atom with an aryl group. Preferred examples thereof include "C5-C10 aryl-C1-C6 alkyl", such as benzyl.

Herein, the term "arylene" means a divalent group derived from the above-mentioned aryl group by removal of another single arbitrary hydrogen atom. The arylene group may be a single ring or a fused ring. The number of ring-forming atoms is not particularly limited, but is preferably 6 to 10(C6-10 arylene). Specific examples of the arylene group include phenylene and naphthylene.

Herein, the term "heteroarylene" means a divalent group derived from the above-mentioned heteroaryl group by removal of another single arbitrary hydrogen atom. The heteroarylene group may be a single ring or a condensed ring. The number of ring-constituting atoms is not particularly limited, but is preferably 5 to 10 (5-to 10-membered heteroarylene). Specific examples of the heteroarylene group include pyrrolediyl, imidazolediyl, pyrazolediyl, pyridinediyl, pyridazinediyl, pyrimidinediyl, pyrazinediyl, triazoldiyl, triazinediyl, isoxazolediyl, oxazolediyl, oxadiazoldiyl, isothiazolediyl, thiazolediyl, thiadiazolediyl, furandiyl, and thiophenediyl.

In the present invention, the "amino acid" constituting the peptide may be a "natural amino acid" or an "amino acid analog". "amino acids", "natural amino acids" and "amino acid analogs" are also referred to as "amino acid residues", "natural amino acid residues" and "amino acid analog residues", respectively.

"Natural amino acid" refers to α -aminocarboxylic acid (α -amino acid), and refers to 20 kinds of amino acids contained in protein.

"amino acid analogs" are not particularly limited and include β -amino acid, γ -amino acid, D-amino acid, N-substituted amino acid, α -disubstituted amino acid, hydroxycarboxylic acid, unnatural amino acid (amino acids whose side chains are different from those of natural amino acids; for example, unnatural α -amino acid, β -amino acid and γ -amino acid). α -amino acid may be D-amino acid, or α -dialkylamino acid in a similar manner to α -amino acid β -amino acid and γ -amino acid are also allowed to have any configuration, the side chains of amino acid analogs (when the main chain is methylene) are not particularly limited and may have, in addition to a hydrogen atom, for example, an alkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, an aralkyl group or a cycloalkyl group each of which may have one or more substituents, and which may be selected from any functional group containing, for example, a halogen atom, an N atom, an O atom, an S atom, a B atom, a Si atom or a P atom, for example, "C-alkoxy group", and "refers to a substituent containing, for example, a halogen atom, such as a substituent, a halogen atom, a substituent such as a halogen atom, a substituent such as a fluorine atom, a substituent such as a substituent, a substituent such as a substituent, a substituent represented by a substituent of 6326, a substituent represented by a substituent of 63.

The backbone amino group of the amino acid analog may be unsubstituted (NH)₂A group) or it may be substituted (i.e., an NHR group; wherein R represents an alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl or cycloalkyl group, each of which optionally has a substituent, or as in proline, the carbon chain bonded to the N atom and the carbon atom at position α may form a ring.

Examples of substituents derived from halogen include fluorine (-F), chlorine (-Cl), bromine (-Br), and iodine (-I).

Examples of the substituent derived from an O atom include a hydroxyl group (-OH), an oxy group (-OR), a carbonyl group (-C ═ O-R), a carboxyl group (-CO)₂H) Oxycarbonyl (-C ═ O-OR), carbonyloxy (-O-C ═ O-R), thiocarbonyl (-C ═ O-SR), carbonylthio (-S-C ═ O-R), aminocarbonyl (-C ═ O-NHR), carbonylamino (-NH-C ═ O-R), oxycarbonylamino (-NH-C ═ O-OR),Sulfonylamino (-NH-SO)₂-R), aminosulfonyl (-SO)₂-NHR), sulfamoylamino (-NH-SO)₂-NHR), thiocarboxyl (-C (═ O) -SH) and carboxycarbonyl (-C (═ O) -CO₂H)。

Examples of the oxy group (-OR) include alkoxy, cycloalkoxy, alkenyloxy, alkynyloxy, aryloxy, heteroaryloxy and aralkyloxy groups.

Examples of the carbonyl group (-C ═ O-R) include formyl group (-C ═ O-H), alkylcarbonyl group, cycloalkylcarbonyl group, alkenylcarbonyl group, alkynylcarbonyl group, arylcarbonyl group, heteroarylcarbonyl group, and aralkylcarbonyl group.

Examples of the oxycarbonyl group (-C ═ O-OR) include an alkoxycarbonyl group, a cycloalkoxycarbonyl group, an alkenyloxycarbonyl group, an alkynyloxycarbonyl group, an aryloxycarbonyl group, a heteroaryloxycarbonyl group, and an aralkoxycarbonyl group.

Examples of carbonyloxy (-O-C ═ O-R) include alkylcarbonyloxy, cycloalkylcarbonyloxy, alkenylcarbonyloxy, alkynylcarbonyloxy, arylcarbonyloxy, heteroarylcarbonyloxy and aralkylcarbonyloxy.

Examples of thiocarbonyl (-C ═ O-SR) include alkylthiocarbonyl, cycloalkylthiocarbonyl, alkenylthiocarbonyl, alkynylthiocarbonyl, arylthiocarbonyl, heteroarylthiocarbonyl and aralkylthiocarbonyl.

Examples of carbonylthio (-S-C ═ O-R) include alkylcarbonylthio, cycloalkylcarbonylthio, alkenylcarbonylthio, alkynylcarbonylthio, arylcarbonylthio, heteroarylcarbonylthio and aralkylcarbonylthio.

Examples of aminocarbonyl (-C ═ O-NHR) include alkylaminocarbonyl, cycloalkylaminocarbonyl, alkenylaminocarbonyl, alkynylaminocarbonyl, arylaminocarbonyl, heteroarylaminocarbonyl and aralkylaminocarbonyl. Further examples include compounds produced by further substitution of the H atom bonded to the N atom in-C ═ O-NHR with an alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl group.

Examples of carbonylamino (-NH-C ═ O-R) include alkylcarbonylamino, cycloalkylcarbonylamino, alkenylcarbonylamino, alkynylcarbonylamino, arylcarbonylamino, heteroarylcarbonylamino and aralkylcarbonylamino. Further examples include compounds produced by further substitution of the H atom bonded to the N atom in-NH-C ═ O-R with an alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl group.

Examples of the oxycarbonylamino (-NH-C ═ O-OR) include alkoxycarbonylamino, cycloalkoxycarbonylamino, alkenyloxycarbonylamino, alkynyloxycarbonylamino, aryloxycarbonylamino, heteroaryloxycarbonylamino and aralkyloxycarbonylamino. Further examples include compounds produced by further substitution of the H atom bonded to the N atom in-NH-C ═ O-OR with an alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, OR aralkyl group.

Sulfonylamino (-NH-SO)₂Examples of-R) include alkylsulfonylamino, cycloalkylsulfonylamino, alkenylsulfonylamino, alkynylsulfonylamino, arylsulfonylamino, heteroarylsulfonylamino and aralkylsulfonylamino. Additional examples include by further substitution of-NH-SO with alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl₂-the H atom of R bonded to the N atom.

Aminosulfonyl (-SO)₂-NHR) include alkylaminosulfonyl, cycloalkylaminosulfonyl, alkenylaminosulfonyl, alkynylaminosulfonyl, arylaminosulfonyl, heteroarylaminosulfonyl and aralkylaminosulfonyl. Additional examples include by further substitution of-SO with alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, or aralkyl₂-H atom bonded to N atom in NHR.

Sulfamoylamino (-NH-SO)₂-NHR) includes alkylsulfamoylamino, cycloalkylsulfamoylamino, alkenylsulfamoylamino, alkynylsulfamoylamino, arylsulfamoylamino, heteroarylsulfamoylamino and aralkylsulfamoylamino. Alternatively, in-NH-SO₂-two H atoms bonded to the N atom in the NHR may be substituted by substituents independently selected from the group consisting of: alkyl, cycloalkyl, alkenyl,Alkynyl, aryl, heteroaryl, and aralkyl; or the two substituents may form a ring.

Examples of the substituent derived from an S atom include a mercapto group (-SH), a thio group (-S-R), a sulfinyl group (-S ═ O-R), a sulfonyl group (-S (O)₂-R) and a sulfo group (-SO)₃H)。

Examples of thio (-S-R) are selected from alkylthio, cycloalkylthio, alkenylthio, alkynylthio, arylthio, heteroarylthio, aralkylthio and the like.

Examples of sulfinyl (-S ═ O-R) include alkylsulfinyl, cycloalkylsulfinyl, alkenylsulfinyl, alkynylsulfinyl, arylsulfinyl, heteroarylsulfinyl and aralkylsulfinyl.

Sulfonyl (-S (O)₂Examples of-R) include alkylsulfonyl, cycloalkylsulfonyl, alkenylsulfonyl, alkynylsulfonyl, arylsulfonyl, heteroarylsulfonyl and aralkylsulfonyl.

For substituents derived from N atoms, examples include azides (-N)₃Also known as "azido"), cyano (-CN), primary amino (-NH)₂) (ii), a secondary amino group (-NH-R), a tertiary amino group (-NR (R')), an amidino group (-C (═ NH) -NH)₂) Substituted amidino (-C (-NR) -NR' R "), guanidino (-NH-C (-NH) -NH)₂) Substituted guanidino (-NR-C (═ NR ' ") -NR ' R") and aminocarbonylamino (-NR-CO-NR ' R ").

Examples of secondary amino groups (-NH-R) include alkylamino, cycloalkylamino, alkenylamino, alkynylamino, arylamino, heteroarylamino and aralkylamino.

Examples of tertiary amino groups (-NR (R')) include amino groups having any two substituents (such as alkyl (aralkyl) amino groups), each independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl; and these two optional substituents may form a ring.

Examples of substituted amidino groups (-C (═ NR) -NR 'R ") include groups in which each of the three substituents R, R' and R" on the N atom are independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl groups; and examples thereof include alkyl (aralkyl) (aryl) amidino groups.

Examples of substituted guanidino (-NR-C (═ NR '") -NR' R") include groups wherein each of R, R ', R ", and R'" is independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl; or groups in which they form a ring.

Examples of aminocarbonylamino (-NR-CO-NR 'R') include those wherein each of R, R 'and R' is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl; or groups in which they form a ring.

Examples of the substituent derived from a B atom include a boryl group (-BR (R ')) and a dioxyboryl group (-B (OR)) (OR')). The two substituents, i.e., R and R', are each independently selected from alkyl, cycloalkyl, alkenyl, alkynyl, aryl, heteroaryl, and aralkyl; or they may form a ring.

In the present invention, at least one atom constituting the "amino acid" may be an atom (isotope) having the same number of atoms (number of protons) and different numbers of masses (total number of protons and neutrons). Examples of isotopes contained in "amino acids" include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms and chlorine atoms, including²H、³H、¹³C、¹⁴C、¹⁵N、¹⁷O、¹⁸O、³¹P、³²P、³⁵S、¹⁸F and³⁶Cl。

exemplary amino acid analogs that can be used in the present invention are described below; however, the amino acid analogs are not limited thereto. Many of these amino acid analogs can be purchased with their side chains protected or unprotected and their amine moieties protected or unprotected. Those that cannot be purchased can be synthesized by known methods.

The following N-Me amino acids can be used as amino acid analogs:

n-methylalanine; n-methylglycine; n-methylphenylalanine; n-methyl tyrosine; n-methyl-3-chlorophenylalanine; n-methyl-4-chlorophenylalanine; n-methyl-4-methoxyphenylalanine; n-methyl-4-thiazolylalanine; n-methyl histidine; n-methyl serine; and N-methyl aspartic acid.

The following N-alkyl amino acids may also be used as amino acid analogs.

The following D-amino acids can also be used as amino acid analogs.

The following α -dialkylamino acids can also be used as amino acid analogs.

The following amino acids may also be used as amino acid analogs.

In one non-limiting embodiment, the present invention provides a process for preparing a cyclic organic compound comprising a cyclization reaction step of cyclizing a cyclization precursor of the cyclic organic compound in at least one continuous stirred tank reactor.

Further, in one non-limiting embodiment, the present invention provides a method of promoting the intramolecular cyclization of a cyclized precursor comprising the step of cyclizing a cyclized precursor of a cyclic organic compound in at least one or more continuous stirred tank reactors.

Additionally, in one non-limiting embodiment, the present invention provides a method of inhibiting intermolecular reactions of cyclized precursors, the method comprising the step of cyclizing a cyclized precursor of a cyclic organic compound in at least one or more continuous stirred tank reactors.

Furthermore, in one non-limiting embodiment, the present invention provides the use of a continuous stirred tank reactor for intramolecular cyclization of a cyclization precursor of a cyclic organic compound in at least one or more continuous stirred tank reactors.

In the present invention, the term "cyclic organic compound" means an organic compound having one or more cyclic moieties. The "cyclic organic compound" in the present invention is not particularly limited as long as it has such a characteristic, but may include, for example, medium molecular weight compounds (e.g., molecular weight of about 500 to 6000), including natural products, sugar chains, peptides and nucleic acid drugs, and low molecular weight compounds (e.g., molecular weight of about 500). Preferred examples include peptide compounds having one or more cyclic moieties.

For example, whether the desired cyclic organic compound has been prepared can be evaluated by measuring the molecular weight of the compound prepared by the method of the present invention using techniques known to those skilled in the art such as MS, SDS-PAGE, and the like.

The term "cyclized precursor" of the present invention means, with respect to the cyclic organic compound of the present invention prepared by performing the cyclization step, a noncyclic organic compound (precursor) before performing the cyclization step. Preferably, but not limitatively, the cyclization precursor has the same chemical structure as the cyclic organic compound except for the structural moieties involved in the cyclization reaction. The moiety involved in the cyclization reaction includes a structure similar to the reaction auxiliary group used for the cyclization reaction, which can be eliminated after the bonding reaction.

For example, when the cyclization precursor is a low molecular weight organic compound, the cyclization precursor can be prepared by using an organic synthesis method, which is a technique known to those skilled in the art. On the other hand, when the cyclized precursor is a peptide compound, it can be obtained by peptide synthesis using a cell-free translation system or expression of a gene encoding the peptide compound in an appropriate host cell, in addition to the above-described organic synthesis method.

In a non-limiting embodiment, the cyclization reaction in the present invention is an intramolecular cyclization reaction through one or more bonds selected from the group consisting of:

(i) an amide bond;

(ii) a disulfide bond;

(iii) an ether bond;

(iv) a thioether bond;

(v) an ester linkage;

(vi) a thioester bond; and

(vii) a carbon-carbon bond.

For example, intramolecular cyclization in the above-described embodiment may be performed by bonding of two amino acids via a disulfide bond, an amide bond, a peptide bond, an alkyl bond, an alkenyl bond, an ester bond, a thioester bond, an ether bond, a thioether bond, a phosphonate ether bond, an azo bond, a C ═ N-C bond, an amide bond, a lactam bridge, a carbamoyl bond, a urea bond, a thiourea bond, an amine bond, a thioamide bond, a sulfinyl bond, a sulfonyl bond, or the like, but the type of bond used for intramolecular cyclization reaction is not limited thereto.

The intramolecular cyclization in the above embodiment is not limited, and is preferably carried out by covalent bonding such as amide bonding, carbon-carbon bonding reaction, S-S bonding, thioether bonding, triazole bonding or benzoxazole bonding (WO 2013/100132; WO 2012/026566; WO 2012/033154; WO 2012/074130; WO 2015/030014; WO 2018/052002; Combchem High through High Screen.2010; 13: 75-87; Nature chem.Bio 2009, 5, 502; Nat Chembiol.2009, 5, 888-90; Bioconjugate chem., 2007, 18, 469-476; ChemBiochem, 2009, 10, 787-798; Chemical Communications (Cambridge, 2011) Uni), 47, 9946-.

Compounds that can be obtained by further chemical modification of the above-mentioned compounds may also be included in the cyclic organic compounds and peptide compounds having one or more cyclic moieties of the present invention.

The peptide compounds of the invention may have one or more linear portions. The number of amide bonds and ester bonds (number/length of natural amino acids or amino acid analogs) is not particularly limited, but when the compound has one or more linear moieties, the total number of residues of the cyclic moiety and the one or more linear moieties is preferably 30 or less. In order to obtain high metabolic stability, the total number of amino acids is more preferably 6 or more, or 9 or more. Further, in addition to the above description, the number of natural amino acids and amino acid analogs constituting the cyclic portion is more preferably 4 to 14, 4 to 13, 5 to 12, 6 to 12, or 7 to 12, and even more preferably 7 to 11 or 8 to 11.9 to 11 residues (10 or 11 residues) are particularly preferred. The number of amino acids and amino acid analogs of the linear portion is preferably 0 to 8, 0 to 7, 0 to 6, 0 to 5, or 0 to 4, and more preferably 0 to 3. The total number of natural amino acid and amino acid analog residues is preferably 6 to 30, 6 to 25, 6 to 20, 7 to 19, 7 to 18, 7 to 17, 7 to 16, 7 to 15, 8 to 14, or 9 to 13. In the present application, unless particularly limited, amino acids include natural amino acids and amino acid analogs.

Furthermore, herein, membrane permeability and metabolic stability means that the peptide compound has sufficient membrane permeability and metabolic stability to be used as a drug, at least when it is used as an oral agent or for targeting an intracellular protein, a nucleic acid, an intracellular domain of a membrane protein, or a transmembrane domain of a membrane protein.

The cyclic moiety of the cyclic moiety-containing peptide compound of the present invention is not particularly limited. As long as they are cyclic, for example, they are preferably cyclic moieties including cyclic moieties formed from functional groups that can satisfy both membrane permeability and metabolic stability.

In cyclization reactions having structurally adjacent reaction sites, such as in 5-membered ring building reactions of indoles and oxirane ring building reactions of epoxides, intramolecular and intermolecular cyclization reactions are less likely to compete with each other (intramolecular cyclization reactions are mainly carried out). Thus, the present invention does not seem to be very advantageous when such a cyclization reaction is used in the present invention.

Herein, "the number of amino acids" refers to the number of amino acid residues (amino acid units) constituting a peptide, and it means the number of amino acid units occurring when the bond linking the amino acids by the amide bond, the ester bond and the cyclized portion is cleaved.

The term "tank reactor" of the present invention refers to a reactor equipped with a stirrer, in which the reaction fluid inside is sufficiently mixed so that the concentration and temperature at any position in the reactor can be considered to be uniform. When the tank Reactor is operated batchwise, it is referred to as a "batch Reactor", and when it is operated continuously, it is referred to as a "continuous stirred tank Reactor" (CSTR) (see "Kagaku Kogaku Binran (Chemical Engineering Handbook), 6 th revision, Maruzen co., ltd., 3.5 bases of Reactor Design). To maximize the productivity, product performance and efficiency of the general process, a single CSTR, a plurality of parallel CSTRs, and a series CSTR in which a plurality of CSTRs are connected in series are used. The reaction raw materials fed into the CSTR were immediately mixed to allow the reaction to proceed, and were discharged from the reactor while maintaining the same concentration and temperature as the inside of the reactor.

In one non-limiting embodiment, the continuous stirred tank reactor used in the present invention can be used not only for homogeneous liquid phase reactions but also for heterogeneous reactions such as liquid-liquid reactions, gas-liquid reactions and gas-liquid-solid catalytic reactions. Furthermore, depending on the type of cyclization reaction of interest and the scale of production, the optimal reactor size can be appropriately selected by those skilled in the art.

In a non-limiting embodiment, in the method of preparing a cyclic organic compound using one or more CSTRs according to the present invention, the CSTRs may be operated together with a reaction tank previously brought into a steady state by being separately performed in a continuous reactor or a batch reactor until a steady state is reached within the reactor or by a reverse dropping method or a high dilution condition method in the batch reactor. Thus, the possibility of discharging unreacted reaction raw materials out of the reactor can be prevented as much as possible, and all the substrates can be used effectively.

In one non-limiting embodiment, a process for preparing cyclic organic compounds using one or more CSTRs is provided, wherein the cyclic organic compounds of the present invention are peptide compounds comprising a cyclic moiety composed of a natural amino acid and/or amino acid analog. Without limitation, the compound preferably comprises a cyclic moiety consisting of a total of 4 to 14 natural amino acid and amino acid analog residues, and has a total of 7 to 20 natural amino acid and amino acid analog residues. Furthermore, the cyclic moiety preferably consists of a total of 5 to 12, 6 to 12, 7 to 12, 8 to 12 or 9 to 12 natural amino acid and amino acid analogue residues, and the total number of natural amino acid analogue residues is preferably 8 to 14, 9 to 13, 10 to 13 or 11 to 13. The total number of amino acids included in the peptide compound is preferably 25 or less, 20 or less, 18 or less, 17 or less, 16 or less, 15 or less, or 14 or less, and more preferably 13 or less (for example, 12, 11, 10, or 9), but is not particularly limited thereto.

In one non-limiting embodiment, the cyclic organic compounds of the present invention have the following characteristics:

(i) comprising at least two N-substituted amino acids and at least one non-N-substituted amino acid; and

(ii) has a ClogP value of 6 or more.

In the above embodiments, the cyclic organic compounds of the invention preferably comprise at least 2 (preferably 2, 3, 4, 5, 6, 7, 8, 9 or 10, and particularly preferably 5, 6 or 7) N-substituted amino acids, and preferably at least 1, 2, 3 or 4 non-N-substituted amino acids. Further, the ClogP value is preferably 7 or more, 8 or more, or 9 or more. "N-substitution" includes substitution of a hydrogen atom bonded to an N atom with a methyl group, an ethyl group, a propyl group, a butyl group, a hexyl group, etc., but is not limited thereto. Preferred N-substituted amino acids include natural amino acids in which the amino group is N-methylated. In the case where the peptide contains a chemically modified amino acid analog, the compound, when considered to have a molecular form (backbone structure) in which all chemical modifications have been completed, preferably has a ClogP value (a distribution coefficient calculated in a computer; for example, it can be calculated using the Daylight version 4.9 of the Daylight chemical Information Systems, inc.) of 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, or 8 or more, and preferably 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, or 15 or less, with respect to the molecule it forms.

The compounds targeted by the cyclization reaction using one or more CSTRs in the present invention are not particularly limited, but their molecular weights are preferably 100 or more, 200 or more, 300 or more, 400 or more, 500 or more, 550 or more, 600 or more, 650 or more, 700 or more, 750 or more, 800 or more, 850 or more, 900 or more, or 950 or more, and preferably 1000 or more, 1100 or more, 1200 or more, 1300 or more, 1400 or more, 1500 or more, 1600 or more, 1700 or more, or 1800 or more. The upper limit of the molecular weight is not particularly limited, but the molecular weight is preferably 6000 or less, 5000 or less, 4000 or less, 3000 or less, 2500 or less, or 2000 or less.

For example, when the value of endogenous liver clearance (CLh int (μ L/min/mg protein)) is 150 or less or preferably 100 or less, 90 or less, 80 or less, 70 or less or 60 or less or particularly preferably 50 or less, 40 or less or 30 or less when measuring stability in liver microsomes according to the above-described method, it can be determined that metabolic stability for use as an oral drug can be obtained. In The case where a drug is metabolized by CYP3A4, The endogenous liver clearance value is preferably 78% or less (non-patent literature: M.Kato et al, The intestinal first-pass metabolism of The substations of CYP3A4 and P-glycoprotein-quantitative analysis of The substance of CYP3A4 and P-glycoprotein quantification based on literature information), drug.Pharmacokinase.2003, 18 (365), and 372) in order to avoid its metabolism in The human small intestine, and is preferably 35% or less (assuming that FaFg is 1 and The protein binding rate is 0%) in order to exhibit bioavailability in humans of about 30% or more.

Further, the cyclic organic compound in the present invention may be a water-insoluble compound. For example, "water-insoluble compound" means a compound having a solubility in ion-exchanged water at 20 ℃ of preferably 10mg/mL or less, or 1mg/mL or less, or more preferably 0.1mg/mL or less, 0.01mg/mL or less, or 0.001mg/mL or less.

In one non-limiting embodiment, the cyclization precursor of the present invention may bear one or more reaction auxiliary groups for the cyclization reaction. In the present invention, the "reaction auxiliary group" means a group which is introduced in the vicinity of the functional group to be bonded and activates the functional group for binding reaction so that the reaction selectively occurs at a desired position. For example, for the reaction between a carbonyl group and an amino group, a reaction auxiliary group may be introduced into the carbonyl side or the amino side or both of them. Examples of such a reaction auxiliary group include SH. Such a reaction auxiliary group may be eliminated simultaneously with or after the binding reaction.

In one non-limiting embodiment, the cyclic organic compounds of the present invention are compounds represented by the following general formula (I):

(wherein X represents CH or N; R1 represents a hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C1-4 haloalkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group, a C1-6 alkoxy group or a C1-6 alkylthio group; R2 represents a hydrogen atom, a halogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group, or R2 and R3 together form a ring; R3 represents a hydrogen atom, a halogen atom, a cyano group, a C1-6 alkyl group, a C2-6 alkenyl group, a C2-6 alkynyl group or a C1-6 alkoxy group, or R2 and R3 together form a ring; R4 represents a hydrogen atom, a C1-6 alkyl group, a C2-6 alkenyl group or a C2-6 alkynyl group; R5, R6 and R39 7 each independently represents an optionally substituted C1-6 alkylene group, a C2-6 alkenylene group, a C2-6 alkenylene group, a C362, C3-10 cycloalkylene, C3-10 cycloalkenylene, C6-12 arylene, or-3 to 12 membered monocyclic heterocycle-; l1, L2 and L3 each independently represent a single bond, -CONR8-, -NR8CO-, -NR8-, -O-, -SO₂NR8-、-NR8SO₂-, -COO-, NR8CONR 8' -, NR8 COO-or-OCONR 8-; and R8 and R8' each independently represent a hydrogen atom or a C1-6 alkyl group which may have a substituent);

the following paragraphs define terms only for the compounds represented by the above general formula (I).

The term "C1-6 alkyl" means a straight or branched chain saturated monovalent C1-6 hydrocarbon group and includes, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, sec-butyl, tert-butyl, 1-methylpropyl, 1-dimethylpropyl, 2, 2-dimethylpropyl, 1, 2-trimethylpropyl, 1, 2, 2-trimethylpropyl, 1, 2, 2-tetramethylpropyl, 1-methylbutyl, 2-methylbutyl, 3-methylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2, 2-dimethylbutyl, 2, 3-dimethylbutyl, 3-dimethylbutyl, 1-ethylbutyl, 2-ethylbutyl, isoamyl and neopentyl.

The term "C1-3 alkyl" means a straight or branched chain saturated monovalent C1-3 hydrocarbon group and includes, for example, methyl, ethyl, propyl, and isopropyl.

The term "C1-4 alkyl" means a straight or branched chain saturated monovalent C1-4 hydrocarbon group and includes, for example, methyl, ethyl, propyl, butyl, isopropyl, sec-butyl, and tert-butyl.

The term "C1-4 haloalkyl" means "C1-4 alkyl" substituted with one or more halogen atoms. Preferably, it is C1-2 alkyl substituted with one or more fluoro or chloro groups and includes, for example, trifluoromethyl, difluoromethyl, fluoromethyl, pentafluoroethyl, tetrafluoroethyl, trifluoroethyl, difluoroethyl, fluoroethyl, trichloromethyl, dichloromethyl, chloromethyl, pentachloroethyl, tetrachloroethyl, trichloroethyl, dichloroethyl and chloroethyl.

The term "C2-6 alkenyl" means a C2-6 hydrocarbon group having at least one double bond, and includes, for example, vinyl (ethenyl), 1-propenyl, 2-propenyl (allyl), isopropenyl, 1-butenyl, 2-butenyl, 3-butenyl (homoallyl), pentenyl and hexenyl.

The term "C2-6 alkynyl" means a C2-6 hydrocarbon group having at least one triple bond, and includes, for example, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, pentynyl, and hexynyl.

The term "C1-6 alkoxy" means-O-C1-6 alkyl, and includes, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, isobutoxy, tert-butoxy, pentyloxy, 3-methylbutyloxy, 2-methylbutyloxy, 1-ethylpropyloxy, hexyloxy, 4-methylpentyloxy, 3-methylpentyloxy, 2-methylpentyloxy, 1-methylpentyloxy, 3-ethylbutoxy and 2-ethylbutoxy.

The term "C1-4 alkoxy" means-O-C1-4 alkyl, and includes, for example, methoxy, ethoxy, propoxy, isopropoxy, butoxy, sec-butoxy, isobutoxy, and tert-butoxy.

The term "C1-3 alkoxy C1-3 alkoxy" means-O-C1-3 alkyl-O-C1-3 alkyl, and includes, for example, methoxymethoxy, methoxyethoxy, and ethoxyethoxy.

The term "C1-6 alkylthio" means-S-C1-6 alkyl, and includes, for example, methylthio, ethylthio, propylthio, isopropylthio, butylthio, sec-butylthio, isobutylthio, tert-butylthio, pentylthio, 3-methylbutylthio, 2-methylbutylthio, 1-ethylpropylthio, hexylthio, 4-methylpentylthio, 3-methylpentylthio, 2-methylpentylthio, 1-methylpentylthio, 3-ethylbutylthio and 2-ethylbutylthio.

The term "halogen" means fluorine (F), chlorine (Cl), bromine (Br) or iodine (I), and is preferably fluorine or chlorine.

The term "C3-10 cycloalkyl" means a saturated C3-10 carbocyclic group and includes, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, and cyclodecyl.

The term "C3-10 cycloalkenyl" means a C3-10 carbocyclic group having at least one double bond, and includes, for example, cyclopentenyl, cyclohexenyl, and cycloheptenyl.

The term "C6-12 aryl" means a monocyclic or bicyclic aromatic carbocyclic ring having 6 to 12 ring carbon atoms, and includes, for example, phenyl, naphthyl, indanyl, indenyl, and isoindolyl. It is preferably phenyl.

The term "3 to 12 membered monocyclic heterocycle" means an aromatic or non-aromatic monocyclic heterocycle having 3 to 12 ring-forming atoms including one or more (e.g., 1 to 4) heteroatoms selected from N, O and S. The bonding position of the heteroatoms is not particularly limited, and they may be bonded at any position. Specific examples of the 3 to 12-membered monocyclic heterocyclic ring include pyrrolidine, oxazolidine, isoxazolidine, oxazoline, isoxazoline, thiazolidine, isothiazolidine, thiazoline, isothiazoline, imidazolidine, imidazoline, furan, thiophene, pyrrole, oxazole, isoxazole, thiazole, isothiazole, furazan, imidazole, pyrazole, piperidine, piperazine, morpholine, thiomorpholine, tetrahydropyran, dioxane, tetrahydrothiopyran, pyran, thiopyran, pyridine, pyrazine, pyrimidine, and pyridazine.

The term "3 to 12 membered monocyclic alicyclic monoaicyclo ring" means a group in which a monocyclic alicyclic hydrocarbon having 3 to 12 ring-forming atoms forms a ring together with one carbon atom on a C1-6 alkylene group. The position of the spiro carbon on the C1-6 alkylene group is not particularly limited, and it may be shared with a desired position. Specifically, the 3 to 12-membered monocyclic alicyclic monoaspirocyclic ring includes, for example, cyclopropanone, cyclobutanone, cyclopentanone, cyclohexanone, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cyclopropene, cyclobutene, cyclopentene and cyclohexene.

The term "3 to 12 membered monocyclic heterocyclic unispiro ring" means a group in which a monocyclic non-aromatic heterocyclic ring having 3 to 12 ring-forming atoms including one or more (e.g., 1 to 4) heteroatoms selected from N, O and S forms a ring together with one carbon atom on C1-6 alkylene. The one or more bonding positions of the one or more hetero atoms are not particularly limited, and they may be bonded at one or more desired positions. Further, the position of the spiro carbon on the C1-6 alkylene group is not particularly limited, and it may be shared by the C1-6 alkylene group at a desired position. Specifically, the 3 to 12-membered monocyclic heterocyclic monoaspiro ring includes, for example, oxetane (oxolane), tetrahydrofuran, tetrahydropyran, thietane (thietane), tetrahydrothiophene, tetrahydrothiopyran, azetidine (azetidine), pyrrolidine, piperidine, oxetanone (oxolone), tetrahydrofuranone, tetrahydropyranone, azetidinone (azetidine), pyrrolidone, piperidone, dioxolane, dioxane, dithiolane (dithiane), and dithiane.

The term "C1-6 alkylene" means a divalent group derived by further removing any hydrogen atom from "C1-6 alkyl" as defined above.

The term "C2-6 alkenylene" means a divalent group derived by further removing any hydrogen atom from "C2-6 alkenyl" as defined above.

The term "C2-6 alkynyl" means a divalent group derived by further removing any hydrogen atom from "C2-6 alkynyl" as defined above.

The term "C3-10 cycloalkylene" means a divalent group derived by further removing any hydrogen atom from "C3-10 cycloalkyl" as defined above.

The term "C3-10 cycloalkenylene" means a divalent group derived by further removing any hydrogen atom from "C3-10 cycloalkenyl" as defined above.

The term "C6-12 arylene" means a divalent group derived by further removing any hydrogen atom from "C6-12 aryl" as defined above.

The term "-3 to 12 membered monocyclic heterocycle-" means a divalent group derived by removing two arbitrary hydrogen atoms from "3 to 12 membered monocyclic heterocycle" as defined above.

When referred to simply as a "ring," the term is meant to encompass all of the aforementioned concepts of "C3-10 cycloalkyl", "C3-10 cycloalkenyl", "C6-12 aryl" and "5-to 12-membered monocyclic heterocycle".

When R5, R6 and R7 in the formula represent "C1-6 alkylene group which may have a substituent", the substituent preferably includes a group selected from group a shown below.

Group A: c1-6 alkyl which may have a substituent (the substituent is hydroxyl or dimethylamino), a halogen atom, hydroxyl, cyano [ R9 and R10 each independently represent a hydrogen atom, C1-3 alkyl or a group represented by-C (═ O) CH 10₃、-C(＝O)CF₃、-C(＝O)CH(NH₂)CH(CH₃)₂or-C (═ O) CH (NH)₂) (4-OH) Ph]A group represented by — C (═ O) NR11R12 [ R11 and R12 each independently represent a hydrogen atom or an optionally substituted C1-6 alkyl group (the substituent represents at least one substituent selected from the group consisting of a hydroxyl group, a C1-6 alkoxy group, a C1-3 alkoxy C1-3 alkoxy group, a morpholinyl group, a piperidinyl group, and a 4-methylpiperidinyl group), or R11 and R12 together form a 3 to 12-membered monocyclic heterocyclic ring]OR a group represented by-C (═ O) OR13 [ R13 represents a hydrogen atom OR C1-3 alkyl group]An optionally substituted 3-to 12-membered monocyclic alicyclic unispiro ring (the substituent represents at least one substituent selected from the group consisting of a hydrogen atom, a halogen atom, a cyano group, a hydroxyl group, a C1-6 alkyl group, a C2-6 alkenyl group, a C1-6 alkoxy group, a C1-6 alkylthio group, a C1-6 acyl group, a carboxyl group, a carbamoyl group, a C1-6 alkoxycarbonyl group, a C1-6 alkoxycarbonyloxy group, a single C1-6 alkylaminocarbonyl group, a single C1-6 alkylaminocarbonyloxy group, a di C1-6 alkylaminocarbonyl group, a di C1-6 alkylaminocarbonyloxy group, an amino group, a single C1-6 alkylamino group, a di C1-6 alkylamino group, a single C1-6 acylamino group, a C1-6 alkylsulfonylamino group, a C1-6 alkoxycarbonylamino group, an N' -single C1-6 ureido group, a, N ', N' -di C1-6 alkylureido, and the like), or an optionally substituted 3 to 12 membered monocyclic heterocyclic unispiro ring (the substituent represents at least one substituent selected from the group consisting of: a hydrogen atom, a halogen atom, a cyano group, a hydroxyl group, an oxy group, a C1-6 alkyl group, a C2-6 alkenyl group, a C1-6 alkoxy group, a C1-6 alkylthio group, a C1-6 acyl group, a carboxyl group, a carbamoyl group, a C1-6 alkoxycarbonyl group, a C1-6 alkoxycarbonyloxy group, a single C1-6 alkylaminocarbonyl group, a single C1-6 alkylaminocarbonyloxy group, a di C1-6 alkylaminocarbonyl group, a di C1-6 alkylaminocarbonyloxy group, an amino group, a single C1-6 alkylamino group, a di C1-6 alkylamino group, a single C1-6 acylamino group, a C1-6 alkylsulfonylamino group, a C1-6 alkoxycarbonylamino group, an N ' -single C1-6 alkylureido group, an N ', N ' -di C1-6 alkylureido group, and the like).

A plurality of such substituents may be present. When a plurality of substituents are present, they may be the same or different. The number of such substituents is preferably 1 or 2.

When R8 and R8' in the formula are "optionally substituted C1-6 alkyl", the substituent preferably includes a group selected from group B shown below.

Group B: hydroxy, morpholinyl, piperidinyl, 4-methylpiperidinyl, a halogen atom, hydroxy, amino, cyano, C6-12 aryl or C1-6 alkoxy.

A plurality of such substituents may be present. When a plurality of substituents are present, they may be the same or different. The number of such substituents is preferably 1 or 2.

< simulation >

In one non-limiting embodiment, the cyclization reaction of the present invention can be carried out on an industrial scale using conditions obtained based on preliminary test results of the cyclization reaction.

In an embodiment of the present invention, the term "simulated" refers to a test that is performed prior to performing an industrial scale reaction (i.e., a process for preparing a cyclic organic compound comprising the step of cyclizing a cyclized precursor of the cyclic organic compound in at least one continuous stirred tank reactor) and is performed in order to obtain optimal conditions for the industrial scale reaction. The simulation of the present invention comprises a step of performing a preliminary test and a step of calculating conditions for performing a cyclization reaction on an industrial scale based on the results of the preliminary test. The "simulation" in the present invention is not particularly limited as long as it is a test conducted for this purpose, and for example, it also includes obtaining conditions already available based on known values and the like described in the literature.

In the present embodiment, "industrial scale" preferably refers to plant-level scale or plant scale, but is not particularly limited thereto. It includes both pilot scale (bench scale) and laboratory scale on the laboratory level, which are carried out at a stage prior to the preparation of the cyclic organic compounds on the plant scale.

Thus, in one non-limiting embodiment, the present invention also provides a process for obtaining conditions for the cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one continuous stirred tank reactor. Furthermore, in one non-limiting embodiment, the present invention also provides a method of obtaining conditions for the preparation of cyclic organic compounds on an industrial scale in at least one continuous stirred tank reactor.

In a preliminary test of the present invention, concentration change data over time at one or more temperatures is obtained for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates, and one or more by-products. Preliminary tests of the present invention can be performed by the methods described in the examples or by methods known to those skilled in the art.

In one non-limiting embodiment, optimal conditions for cyclization on an industrial scale using one or more CSTRs can be obtained by a process comprising the steps of:

(i) in the preliminary test, concentration change data with time at a plurality of temperatures was obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) determining the reaction rate constant k for each temperature subjected to the preliminary test by using the concentration change data obtained in step (i) and the reaction rate equations for the cyclization reaction (equations (V) to (IX) or (XI) to (XIII) herein)_n；

(iii) (iii) by using the temperature used in step (i), the reaction rate constant k determined in step (ii)_nAnd the following equation (II), determining the frequency factor A_nAnd activation energy E_n：

(where k is_nDenotes the reaction rate constant, A_nDenotes a frequency factor, E_nRepresents activation energy, R represents a gas constant, and T represents temperature);

(iv) by using the frequency factor A determined in step (iii)_nAnd activation energy E_nEquation (II) above and the reaction rate equation above, determine the reaction rate constant k at the cyclization temperature in one or more CSTRs_n(ii) a And

(v) (iii) by using the reaction rate constant k determined in step (iv)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

In the above embodiment, the "plurality of temperatures" of step (i) is preferably 2 or more, 3 or more, 4 or more, or 5 or more temperatures, but is not limited thereto.

In the above embodiment, the reaction rate constant (k) at the cyclization temperature in the CSTR can be determined by substituting the frequency factor value (a) and the activation energy value (E) obtained in step (III) into the above Arrhenius equation (Arrhenius' equalisation) (III). By substituting the reaction rate constant (k) at the cyclization temperature in the CSTR into the reaction rate equation of the cyclization reaction and setting the initial concentration conditions, the concentration change of each component with time at the cyclization temperature in the CSTR can be determined. In the present invention, the cyclization temperature in the CSTR can be set arbitrarily, which allows optimization of the temperature for industrial scale cyclization.

In the present invention, the optimum condition of "cyclization temperature in CSTR" is preferably a temperature condition of cyclization reaction in CSTR when the concentration of the target molecule (concentration of cyclic organic compound) is maximized, but is not limited thereto.

Carrying out the above-mentioned steps (i) to (v) makes it possible to simulate the progress of the cyclization reaction at any temperature (variation in the concentration of the components with time), and thus to predict, for example, the reaction conversion (percentage of cyclized precursor used for the reaction) and selectivity (percentage of target molecule in all the products) in the reaction. Based on the results of such a simulation, optimum conditions (for example, supply concentration (initial concentration) of the cyclization precursor, and residence time in the reactor, reaction temperature, etc.) for the reaction on an industrial scale can be determined.

In one non-limiting embodiment, one skilled in the art may also suitably select the cyclization temperature in one or more CSTRs of step (iv) from among the temperatures used in step (i) to obtain concentration variation data or from temperatures in the vicinity of these temperatures (+ -5 ℃ or + -10 ℃) without performing the step of optimizing the temperature on a commercial scale described above.

In one non-limiting embodiment, optimal conditions for industrial scale cyclization using one or more CSTRs can be obtained by a process comprising the steps of:

(i) in the preliminary test, concentration change data with time at a single temperature was obtained for at least one selected from the group consisting of: a cyclization precursor, a cyclic organic compound, one or more intermediates and one or more side products;

(ii) (ii) determining a reaction rate constant k by using the concentration change data obtained in step (i) and reaction rate equations (V) to (IX) or equations (XI) to (XIII) below) relating to the cyclization reaction_n(ii) a And

(iii) (iii) use of the reaction rate constant k determined in step (ii)_nThe above conditions were obtained with the above reaction rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

In the present invention, "one or more intermediates" refers to one or more species generated during a chemical reaction of a reactant to a product. When the chemical reaction is a multi-step reaction, it refers to a substance generated in each elementary reaction.

In the present invention, "one or more by-products" refers to one or more products other than the main product (target molecule) among the products produced by the chemical reaction. The by-products in the present invention include multimers (linear forms) of cyclized precursors of cyclic organic compounds, cyclized forms of these multimers, and the like, but are not limited thereto. Multimers include dimers, trimers, and the like, but are not limited thereto.

In the present invention, "concentration change" refers to a change in concentration of a substance to be measured with respect to time. In the present invention, it is necessary to obtain data on the change in concentration of the cyclized precursor. In addition, it is preferable to obtain the data of the change in concentration of at least one selected from the group consisting of the cyclic organic compound, one or more intermediates, and one or more by-products. In particular, in the present invention, it is preferable to obtain data on the change in concentration of the cyclization precursor and the cyclic organic compound, and it is particularly preferable to obtain data on the change in concentration of the cyclization precursor, the cyclic organic compound, and one or more by-products. Concentration variation data can be obtained by methods known to those skilled in the art, such as the methods described in the examples.

In one non-limiting embodiment, the motif of the cyclization reaction of the present invention can be represented by the following formula (IV):

here, in the above (IV), when the reaction to produce the Intermediate (IM) from the cyclization precursor (SM) and the Activator (ACT) is faster than the reaction to produce the Target Molecule (TM) and the Dimer (Dimer) from the Intermediate (IM), the motif reaction of the cyclization reaction of the present invention can also be represented by the following formula (X) in one non-limiting embodiment:

when the elementary reaction of the cyclization reaction is represented by the above formula (IV), the reaction rate constant k₁、k₂And k₃Can be determined using any one or a combination of the following equations (V) to (IX):

(wherein r represents a reaction rate, TM represents a cyclic organic compound, SM represents a cyclized precursor, ACT represents an activator, IM represents an intermediate, Dimer represents a Dimer, and C represents a concentration (M)).

K above₁、k₂And k₃Corresponding to the reaction rate constants in the following reactions in the above formula (IV):

k₁: cyclized precursor (SM) + Activator (ACT) → Intermediate (IM)

k₂: intermediate (IM) → Target Molecule (TM)

k₃: intermediate (IM) → Dimer (Dimer)

On the other hand, when the elementary reaction of the cyclization reaction is represented by the above formula (X), the reaction rate constant k₁And k₂Can be determined using any one or a combination of the following equations (XI) through (XIII):

(where r denotes reaction rate, TM denotes target molecule, SM denotes cyclized precursor (═ intermediate), ACT denotes activator, Dimer denotes Dimer, and C denotes concentration (M)).

K above₁And k₂Corresponding to the reaction rate constants in the following reactions in the above formula (X):

k₁: cyclized precursor (SM) (═ Intermediate (IM)) (+ Activator (ACT)) → Target Molecule (TM)

k₂: cyclized precursor (SM) (═ Intermediate (IM)) (+ Activator (ACT)) → Dimer (Dimer)

The person skilled in the art can appropriately determine whether the simplified reaction formula is applicable as in the above formula (X) according to the accuracy of the experimental data required. In general, those skilled in the art can appropriately judge it according to whether or not the concentration change results with time obtained in advance for the above cyclic organic compounds can be reproduced using the following equations (XI) to (XIII). As a criterion for judgment, a residual sum of squares (residual sum of squares) may be used.

Further, in the case where the concentration of the Activator (ACT) is higher than the concentration of the cyclized precursor (SM) such as in the above-mentioned formulas (IV) and (X), the concentration of the activator (C) in the reaction rate equations (V) to (IX) and (XI) to (XIII)_ACT) And in some cases may be omitted. The skilled person can appropriately judge whether such omission is applicable or not according to the accuracy of the experimental data required. In general, the skilled person can omit the activator concentration (C) depending on whether it is possible to use_ACT) The above-mentioned cyclic organic compound is appropriately judged by reproducing the concentration change result with time obtained in advance for the above-mentioned cyclic organic compound. As a criterion for judgment, the sum of squares of the residuals may be used.

Alternatively, in the present invention, the above-described rate constant electricity may be calculated as follows. By taking the logarithm, the above arrhenius equation (II) can be expressed as the following equation (XIV):

the results of the concentration change with time of the above cyclic organic compound obtained by cyclizing the above cyclized precursor and the above reaction rate equations (V) to (IX) and (XI) to (XIII) are used to calculate the reaction rate constant (k) in these equations. The logarithm of the reaction rate constant (lnk) and the reciprocal of the temperature at which cyclization was carried out (1/T) were plotted on the ordinate and abscissa, respectively, to generate an Arrhenius curve. The frequency factor value (A) and the activation energy value (E) can then be determined by using regression analysis techniques known to those skilled in the art (intercept: lnA; and slope: -E/R).

Those skilled in the art can suitably perform these calculations using, for example, Excel supplied by Microsoft Corporation (Microsoft Corporation). On the other hand, in addition to the above-described techniques, the frequency factor value (a) and the activation energy value (E) can be directly determined using software. For example, gpromes provided by PSE and Aspen Batch model provided by Aspen Technology may be used.

By substituting the frequency factor value (a) and the activation energy value (E) obtained via the above-described technique into the above-described arrhenius equation (II), the reaction rate constant (k) at any temperature can be determined. By substituting the reaction rate constant (k) at an arbitrary cyclization temperature in the CSTR into the reaction rate equations (V) to (IX) and (XI) to (XIII) and by setting the initial concentration conditions, the concentration change of each component with time at an arbitrary temperature can be determined. Determining the concentration change over time enables optimization of the temperature used in industrial scale operations. The optimal condition of the "cyclization temperature in CSTR" of the present invention is preferably a cyclization reaction temperature condition that will maximize the concentration of the target molecule, but is not limited thereto.

In the present invention, the optimum values of the cyclization reaction conditions for industrial scale operations can be calculated by using:

1) reaction rate constant k determined by the method described above_nOr is or

2) Reaction rate constant k determined by using the concentration change data obtained at a single temperature and the reaction rate equations (V) to (IX) and (XI) to (XIII) relating to the cyclization reaction_nAnd are and

the following CSTR mass balance equation (XV):

(wherein r is_nThe reaction rates ((V) to (IX) and (XI) to (XIII)) are shown, τ is the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

For example, the concentration of each component at the outlet can be determined by setting the residence time, supply concentration, and temperature and using equation (XV) and equations (V) to (1X) and (XI) to (XIII). When a simple reaction is carried out in a CSTR, the mass balance of the reactant A allows the cumulative amount (accumulation term) to be 0 when assuming a steady state, which can be expressed by the following equation (see Kagaku Kogaku (Chemical Engineering) (3 rd revised edition) Kaisetsu to Enshu (interpretation and Practice)) (edited and supervised by the society of Chemical Engineers of Japan; Y., Tada edition) Chapter 12.5 continuous stirred tank reactor (page 316)):

v₀C_A，0-vC_A-(-r_A)V＝0 (XVI)

(wherein v is₀Denotes the volume of the influent per unit time of the reaction system, C_A，0Denotes the concentration of A in the influent (initial concentration), v denotes the volume of effluent per unit time of the reaction system, C_ARepresents the concentration of A in the effluent (outlet concentration), r_AThe reaction rate of A (change in concentration of A per unit time), V the reactor volume, V₀C_A，0Denotes the inflow per unit time A, vC_ARepresents the outflow per unit time A, (-r)_A) V represents the reaction amount per unit time a, and 0 on the right side represents the cumulative amount).

When the reaction system is a liquid, and assuming that the volume change accompanying the reaction is negligible, the flow rate can be considered as v ═ v₀. Then,. tau. ═ V (reactor volume)/V was used₀Conversion of equation (XVI) (flow per unit time) yields the following equation, referred to as CSTR, which relates to the equation:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Denotes the supply concentration, C denotes the concentration, and X_AIndicating the reaction conversion).

τ ═ V (reactor volume)/V₀The (flow per unit time) represents how large the reactor volume is as much as the volume processed per unit time, or the time it takes to process a feedstock equal to the reactor volume, and is called space time. The inverse space-time is called the Space Velocity (SV).

In one non-limiting embodiment, the optimal cyclization reaction condition values on an industrial scale in the present invention are one or more reaction condition values selected from the group consisting of: the flow rate of the continuous stirred tank reactor, the concentration of the cyclization precursor, and the concentration of the cyclic organic compound, but are not limited thereto.

Furthermore, in one non-limiting embodiment, the present invention relates to a program for causing a computer to perform the following steps to obtain conditions for cyclizing a cyclization precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR) on an industrial scale, and to a recording medium having such a program recorded therein:

(i) for determining the reaction rate constant k at each temperature for preliminary testing by using_nThe steps of (1):

data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates and one or more by-products, and

one or more reaction rate equations (V) to (IX) or (XI) to (XIII) herein) relating to the cyclization reaction;

(ii) (ii) a reaction rate constant k determined in step (i) by using the temperature used in step (i)_nAnd equation (II) below to determine frequencyFactor A_nAnd activation energy E_nThe process of (a):

(where k is_nDenotes the reaction rate constant, A_nDenotes a frequency factor, E_nRepresents activation energy, R represents a gas constant, and T represents temperature);

(iii) for use by using the frequency factor A determined in (ii)_nAnd activation energy E_nThe reaction rate constant k at the cyclization temperature in the CSTR is determined by equation (II) above and one or more of the reaction rate equations above_nA step (2); and

(iv) for using the reaction rate constant k determined in (iii)_nOne or more of the above reaction rate equations and the following CSTR mass balance equation (III) to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

In one non-limiting embodiment, the program of the present invention is a program for causing a computer to execute the steps of:

(i) for determining the reaction rate constant k by using_nThe steps of (1):

concentration change data over time at a single temperature for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates and one or more by-products, and

one or more reaction rate equations (V) to (IX) or (XI) to (XIII) herein) relating to the cyclization reaction; and

(ii) for using the reaction rate constant k determined in (i)_nOne or more of the above-mentioned reactionsThe steps to achieve the above conditions should be taken with the rate equation and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

In one non-limiting embodiment, the program of the present invention is a program for performing the above steps using a computer comprising means for inputting data, means for storing data, means for processing data, and means for outputting data. The procedure of the present invention can be obtained by a method known to those skilled in the art based on the formulae (IV) and (X) herein, which are elementary reaction formulae of cyclization reaction, the equations (V) to (IX) and (XI) to (XIII) for calculating reaction rate constants, the arrhenius equation (II), the CSTR mass balance equation (III), and the like.

Furthermore, in one non-limiting embodiment, the present invention provides a system for obtaining conditions for cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR), the system comprising:

(i) the reaction rate constant k at each temperature for preliminary test was determined by using_nThe device of (2):

data obtained in preliminary tests on the change in concentration over time at a plurality of temperatures for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates and one or more by-products, and

one or more reaction rate equations (V) to (IX) or (XI) to (XIII) herein) relating to the cyclization reaction;

(ii) (ii) by using the temperature used in device (i), the reaction rate constant k determined in step (i)_nAnd the following equation (II) to determine the frequency factor A_nAnd activation energy E_nThe device of (2):

(where k is_nDenotes the reaction rate constant, A_nDenotes a frequency factor, E_nRepresents activation energy, R represents a gas constant, and T represents temperature);

(iii) by using the frequency factor A determined in step (ii)_nAnd activation energy E_nThe reaction rate constant k at the cyclization temperature in the CSTR is determined by equation (II) above and one or more of the reaction rate equations above_nThe apparatus of (1); and

(iv) (iv) use of the reaction rate constant k determined by device (iii)_nThe above reaction rate equation and the following CSTR mass balance equation (III) to obtain the above conditions:

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

Furthermore, in one non-limiting embodiment, the present invention provides a system for obtaining conditions for cyclization on an industrial scale of a cyclized precursor of a cyclic organic compound in at least one Continuous Stirred Tank Reactor (CSTR), the system comprising:

(i) the reaction rate constant k was determined by using_nThe device of (2):

data obtained in preliminary tests on the change in concentration over time at a single temperature for at least one selected from the group consisting of: a cyclic precursor, a cyclic organic compound, one or more intermediates and one or more by-products, and

one or more reaction rate equations (V) to (IX) or (XI) to (XIII) herein) relating to the cyclization reaction; and

(ii) using the reaction rate constant k determined by device (i)_nOne orMeans for deriving said conditions from a plurality of said reaction rate equations and the following CSTR mass balance equation (III):

(wherein r is_nDenotes the reaction rate,. tau.denotes the residence time (space time), C₀Representing the supply concentration, and C representing the concentration).

< pharmaceutical composition >

The present invention provides peptide compounds prepared by the methods of the invention, as well as pharmaceutical compositions comprising such compounds.

In addition to the peptide compounds prepared by the method of the present invention, the pharmaceutical compositions of the present invention can be formulated by known methods by introducing a pharmaceutically acceptable carrier. Conventional excipients, binders, lubricants, colorants, flavors and, if necessary, stabilizers, emulsifiers, absorption promoters, surfactants, pH adjusters, preservatives, antioxidants and the like may be used for formulation, and they are mixed with ingredients generally used for raw materials of pharmaceutical preparations and formulated by a conventional method.

For example, an oral formulation is prepared by: excipients and, if necessary, binders, disintegrants, lubricants, colorants, flavors, and the like are added to the compound according to the present invention or a pharmaceutically acceptable salt thereof, and then they are formulated into powders, fine granules, tablets, coated tablets, capsules, and the like by a conventional method.

Examples of such ingredients include: animal oils and vegetable oils such as soybean oil, beef tallow, and synthetic glycerides; hydrocarbons such as liquid paraffin, squalane and paraffin wax; ester oils such as octyldodecyl myristate and isopropyl myristate; higher alcohols such as cetostearyl alcohol and behenyl alcohol; a silicone resin; a silicone oil; surfactants such as polyoxyethylene fatty acid esters, sorbitan fatty acid esters, glycerin fatty acid esters, polyoxyethylene sorbitan fatty acid esters, polyoxyethylene hydrogenated castor oil, and polyoxyethylene-polyoxypropylene block copolymers; water-soluble polymers such as hydroxyethyl cellulose, polyacrylic acid, carboxyvinyl polymers, polyethylene glycol, polyvinylpyrrolidone, and methyl cellulose; lower alcohols such as ethanol and isopropanol; polyols such as glycerol, propylene glycol, dipropylene glycol, and sorbitol; sugars such as glucose and sucrose; inorganic powders such as silicic anhydride, magnesium aluminum silicate, and aluminum silicate; and purified water.

Examples of excipients include lactose, corn starch, white soft candy, glucose, mannitol, sorbitol, microcrystalline cellulose, and silicon dioxide.

Examples of the binder include polyvinyl alcohol, polyvinyl ether, methyl cellulose, ethyl cellulose, gum arabic, tragacanth, gelatin, shellac, hydroxypropylmethyl cellulose, hydroxypropylcellulose, polyvinylpyrrolidone, polypropylene glycol-polyoxyethylene block polymer, and meglumine.

Examples of disintegrants include starch, agar, gelatin powder, microcrystalline cellulose, calcium carbonate, sodium bicarbonate, calcium citrate, dextrin, pectin, and carboxymethylcellulose calcium.

Examples of lubricants include magnesium stearate, talc, polyethylene glycol, silicon dioxide and hydrogenated vegetable oils.

The colorants used are those approved for use as pharmaceutical additives. The correctant is cocoa powder, Mentholum, empasm, oleum Menthae Dementholatum, Borneolum Syntheticum, cortex Cinnamomi Japonici powder, etc.

It will be apparent that these tablets and granules may be sugar-coated or otherwise coated as appropriate, if desired. Liquid preparations such as syrup and injectable preparations are prepared by adding a pH adjuster, a solubilizer, a tonicity adjuster or the like and, if necessary, a solubilizer, a stabilizer or the like to the compound according to the present invention or a pharmaceutically acceptable salt thereof, and are formulated by a conventional method.

For example, they may be used parenterally in the form of injections, either in sterile solutions or suspensions with water or other pharmaceutically acceptable liquids. For example, they may be formulated in the following manner: they are suitably combined with a pharmaceutically acceptable carrier or medium, particularly with sterile water, physiological saline, vegetable oils, emulsifiers, suspensions, surfactants, stabilizers, flavoring agents, excipients, vehicles, preservatives, binders and the like, and mixed in unit dosage forms as required by generally accepted pharmaceutical practice. Specifically, the carrier includes light anhydrous silicic acid, lactose, crystalline cellulose, mannitol, starch, carboxymethylcellulose calcium, carboxymethylcellulose sodium, hydroxypropylcellulose, hydroxypropylmethylcellulose, polyvinyl acetal diethylaminoacetate, polyvinylpyrrolidone, gelatin, medium chain triglyceride, polyoxyethylene hydrogenated castor oil 60, sucrose, carboxymethylcellulose, corn starch, inorganic salts, and the like. In such formulations, the amount of active ingredient should be such that it is in an appropriate amount within the specified range.

Sterile compositions for injection can be formulated using vehicles such as distilled water for injection in accordance with standard pharmaceutical practice.

Aqueous solutions for injection include, for example, physiological saline and isotonic solutions containing glucose or other adjuvants such as D-sorbitol, D-mannose, D-mannitol, and sodium chloride. Suitable solubilizers may also be used in combination, for example alcohols, particularly ethanol, polyols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants (e.g., polysorbate 80 (registered trademark), HCO-50).

The oily liquid includes sesame oil and soybean oil, and they may be used in combination with benzyl benzoate and/or benzyl alcohol as a solubilizing agent. Buffers (e.g., phosphate buffers and sodium acetate buffers), analgesics (e.g., procaine hydrochloride), stabilizers (e.g., benzyl alcohol and phenol), and/or antioxidants may also be combined. Typically, the prepared injectate is filled into appropriate ampoules.

Administration is preferably oral administration, but is not particularly limited to oral administration. Parenteral administration specifically includes injection, nasal administration, pulmonary administration or transdermal administration. The injection can be administered systemically or locally, for example, by intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection, etc.

The appropriate method of administration may be selected according to the age and symptoms of the patient. The dosage of the pharmaceutical composition comprising the peptide compound prepared by the method of the present invention may be selected in the range of, for example, 0.0001mg to 1000mg per 1kg body weight per administration. Alternatively, the dose may be selected, for example, in the range of 0.001mg to 100000mg per patient; however, the dosage is not necessarily limited to these values. Although the dose and administration method are different depending on the body weight, age, symptoms, and the like of the patient, one skilled in the art can select an appropriate dose and administration method.

All prior art documents cited herein are incorporated by reference into this specification.

Examples

Hereinafter, suitable specific embodiments of the present invention will be described with reference to examples, but should not be construed as being limited thereto.

^2，6[ example 1]4-amino-18, 20-dimethyl-7-thia-3, 5, 11, 15-tetraaza-tricyclo [15.3.1.1] Synthesis of docodecane-1 (20), 2(22), 3, 5, 17(21), 18-hexene-10, 16-dione

The scheme is as follows:

simulation of reaction Rate parameters

Concentration change data for each compound as a function of temperature/reaction time was collected to calculate a reaction rate parameter. The cyclization precursor 2 mesylate monohydrate (7mM) prepared in a 1: 1 solution of N, N-Dimethylacetamide (DMA) and acetonitrile (solution A) and O- (benzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HBTU) and Diisopropylethylamine (DIPEA) prepared in a 1: 1 solution of DMA and acetonitrile containing concentrations of 10mM and 32mM, respectively (solution B) were reacted at 0 deg.C, 24 deg.C or-9 deg.C. The reaction was quenched with methylamine solution. The reaction was carried out using a plug flow reactor, and the reaction time was adjusted by the flow rate of the solutions a and B to obtain the data shown in table 1.

The compound concentration was obtained by assuming that the molar absorbances of the cyclization precursor and the target molecule were the same, and converting the initial concentration of the cyclization precursor (3.5mM, as the reaction mixture solution of the above experiment) to HPLC area%.

[ Table 1]

Data on the change in chemical concentration (example 1)

From the data obtained, the mechanism of this cyclization reaction is considered as follows. The inventors hypothesized that the reaction to generate reactive intermediates from cyclized precursors and HBTU is relatively fast, and that the reaction to generate target molecules and dimers from reactive intermediates is the rate limiting step (James c. collins and Keith James, med. chem. commun., 2012(3), 12, 1489-. In addition, since the activator concentration does not have a great influence on the reaction rate, it is omitted. On the other hand, some reactive intermediate remained and was detected as by-product 1 when the reaction was quenched with methylamine. The sum of byproduct 1 and starting material was taken as the starting material concentration and used to calculate each parameter for subsequent experiments.

Byproduct 1: the N-formamide (N-methyl amide) product; LCMS: ESI (m/z): calculated values: 417.21, found: 417[ M + H ]]⁺。

Using the data on the change of concentration with time obtained by the experiment and the temperature data (Table 1), the reaction rate constants k1 and k2 at each temperature (0 ℃, 24 ℃ and-9 ℃) for each elementary reaction were obtained. The reaction rate constants k1 and k2 were calculated using the following equations:

SM: cyclizing the precursor; TM: a target molecule; dimer: dimer (linear); c-Dimer: dimers (cyclic); c: a concentration (M);k_n: constant of reaction rate

The frequency factor (a) and activation energy (E) of each elementary reaction were obtained from the arrhenius curve according to the resulting reaction rate constants k1 and k2 at each temperature and according to the temperature. The calculation was performed using Microsoft Excel.

As a result, the data shown in table 2 were obtained.

[ Table 2]

Frequency factor (A) and activation energy (E) corresponding to reaction rate constants k1 and k2

	A	E[kJ/mol]
			k1	1.387E+06[1/sec]	43.17
k2	3.513E+08[L/mol/sec]	46.09

The obtained frequency factor (a) and activation energy (E) of each elementary reaction were substituted into the following arrhenius equation, and by using the above reaction rate equation, reaction rate constants k1 and k2 were calculated in the case where the temperature at which the cyclization reaction was carried out in the CSTR was set to 0 ℃ (table 3).

[ Table 3]

Reaction rate constants k1 and k2 for carrying out the cyclization reaction in a CSTR

Temperature of	k1	k2
				0℃	7.69E-03	5.38E-01

Next, using the reaction rate constants k1 and k2 of table 3, the following mass balance equation and reaction rate equation, the reaction conversion, selectivity and residence time when the cyclized precursor concentration was set to 0.05mol/L and the reaction temperature was set to 0 ℃ were calculated (table 4).

[ Table 4]

Calculated reaction conditions (when the concentration of the cyclized precursor was 0.05mmol/L and the reaction temperature was 0 ℃ C.)

	Conversion rate of reaction	Selectivity	Retention time [ theta, min ]]
				Case 1	0.98	0.93	89
Case 2	0.99	0.96	165
				Case 3	0.99	0.98	330

In cases 1 to 3 of Table 4, the residence time that will produce the results that will make the conversion and selectivity of the reaction higher was determined by calculation. Case 2 (run 4) and case 3 (run 5) were performed in this example.

Calculation formula

Reaction conversion rate: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

And (3) selectivity: target molecule concentration/(target molecule concentration + dimer (straight chain + cyclic) concentration)

Run 1: batch high dilution

According to the patent document (JP 5235859) and the non-patent document (A. Suda et al, Bioorganic & Medicinal Chemistry Letters 22(2012)1136-1141), the cyclized precursor 2 methanesulfonate monohydrate (39.9mg, 86% content, 0.056mmol) was dissolved in N, N-Dimethylformamide (DMF) (16.3mL) and tetrahydrofuran (16.3mL), and N-hydroxybenzotriazole (HOBt) monohydrate (50.2mg, 5.9 equivalents), DIPEA (0.23mL, 23.6 equivalents) and 1-ethyl-3- (3' -dimethylaminopropyl) carbodiimide hydrochloride (125.7mg, 11.7 equivalents) were added to the mixture in this order. The resulting mixture was allowed to react at room temperature for 13 hours. When the solution concentration of the target molecule in the reaction solution was quantified to calculate the yield, it corresponded to 18.5mg (86% yield). (according to A.Suda et al, Bioorganic & Medicinal Chemistry Letters 22(2012)1136-1141, the two-step isolation yield is 21%).

Run 2: batch high dilution

Following run 1, the cyclized precursor (40.5mg, 86% content, 0.057mmol) was dissolved in DMF (16.3mL) and acetonitrile (16.3mL), and HBTU (30.5mg, 1.4 equiv.) and DIPEA (0.046mL, 4.6 equiv.) were added to the mixture in that order. The resulting mixture was allowed to react at room temperature for 1 hour. When the solution concentration of the target molecule in the reaction solution was quantified to calculate the yield, it corresponded to 18.4mg (84% yield).

Run 3: batch pseudo high dilution

To a solution of HBTU (0.7441g, 1.4 eq) and DMA (5mL) in acetonitrile (15mL) was added dropwise a solution of the cyclized precursor compound (1.0036g, 86% content, 1.41mmol) and DIPEA (1.14mL, 4.6 eq) in DMA (10mL) at 0 ℃ over a period of 300 minutes. After 60 minutes, when the concentration of the solution of the target molecule was quantified to calculate the yield, it corresponded to 485.2mg (89% yield).

Run 4: CSTR (continuous stirred tank reactor)

The cyclized precursor (2.3004g, 86% content, 3.22mmol) and DIPEA (2.6mL, 4.6 equiv.) were dissolved in DMA (31.9 mL). The substrate concentration was 0.09 mmol/mL. Meanwhile, HBTU (1.7042g, 1.4 eq) was dissolved in acetonitrile (34.5 mL). The reagent concentration was 0.13 mmol/mL. These solutions were added to 16.5mL of each of the DMA/acetonitrile (1/1) solutions at 0 ℃ at an input rate of 0.05mL/min, and the reaction solutions were simultaneously withdrawn at an output rate of 0.1 mL/min. Samples were taken by collecting the reaction solutions 165 minutes (θ), 330 minutes (2 θ), 495 minutes (3 θ) and 660 minutes (4 θ) after the start of the operation, and the samples were quenched using dimethylamine solution. All output solutions were put together and when quantified to calculate the yield, it corresponded to 987.2mg (87% yield).

Theta is elapsed time/mean residence time

Run 5: CSTR (continuous stirred tank reactor)

The cyclized precursor (2.3001g, 86% content, 3.22mmol) and DIPEA (2.6mL, 4.6 equiv.) were dissolved in DMA (31.9 mL). The substrate concentration was 0.09 mmol/mL. Meanwhile, HBTU (1.7083g, 1.4 eq) was dissolved in acetonitrile (34.5 mL). The reagent concentration was 0.13 mmol/mL. These solutions were added to 33mL of each of the DMA/acetonitrile (1/1) solutions at 0 ℃ at an input rate of 0.05mL/min, and the reaction solutions were simultaneously withdrawn at an output rate of 0.1 mL/min. Samples were taken by collecting the reaction solution 165 minutes (0.5 θ), 330 minutes (θ) and 495 minutes (1.5 θ) after the start of the operation, and the samples were quenched using dimethylamine solution.

Analysis conditions were as follows:

HPLC, LCMS conditions:

column: ascentis C18, 4.6mm I.D.. times.50 mmL, 2.7 μm, Supelco

Mobile phase: A) water to TFA 2000: 1, B) acetonitrile to TFA 2000: 1

Column temperature: 30 deg.C

Flow rate: 1.0mL/min

Gradient (B%): 0-3.5min (5 → 23), 3.5-8.0min (23 → 61), 8.0-9.5min (61 → 100), 9.5-9.6min (100), 9.6-9.7min (100 → 5), 9.7-12.0min (5).

MS detection mode: ESI (LC/MS): m/z

Target molecule:

LCMS：ESI(m/z)：386[M+H]⁺and (3) running: 3.9min

¹H NMR (DMSO-d 6): as in the patent literature

Cyclization precursor:

the content is as follows: 86% (from)¹Data on HMNMR: DMSO-d6, internal standard: 1, 3, 5-trimethoxybenzene)

LCMS: ESI (m/z): calculating a value;404.18, found value; 404[ M + H]⁺Elution time: 3.6min

¹H-NMR(DMSO-D₆)δ：8.14(1H，t，J＝5.7Hz)，7.91(1H，s)，7.76-7.66(1H，m)，7.73(2H，br s)，7.34(1H，s)，6.99(1H，s)，3.41(2H，t，J＝6.8Hz)，3.16-3.12(2H，m)，2.83-2.77(2H，m)，2.60(2H，t，J＝6.8Hz)，2.57(3H，s)，2.40(6H，s)，2.38(3H，s)，1.73-1.67(2H，m)。

Dimer (cyclized form)

LCMS: ESI (m/z): calculated values: 771.32, found: 771[ M + H]⁺Elution time: 4.7min

Dimer (straight chain form)

LCMS: ESI (m/z): calculated values: 789.33, found: 789[ M + H ]]⁺Elution time: 4.6min

As a result:

[ Table 5]

Results of reaction purity in runs 1-5: HPLC area% (230nm)

	Cyclized precursors	Target molecules	Dimer (cyclized form)	Dimer (straight chain form)
					Operation 1	N.D.	93.229	5.754	N.D.
Operation 2	N.D.	90.057	5.100	N.D.
					Run 3	N.D.	95.706	3.336	N.D.
Run 4, after reaction	N.D.	94.241	5.400	N.D.
					Run 5, 495nm (1.5 θ)	0.353	96.429	3.107	N.D.

[ Table 6]

Time course of reaction purity in run 4: HPLC area% (230nm)

Time after starting operation	Cyclized precursors	Target molecules	Dimer (cyclized form)	Dimer (straight chain form)
					165 minutes (theta)	2.034	90.737	6.211	N.D.
330 minutes (2 theta)	1.501	91.792	5.961	N.D.
					495 minutes (3 theta)	1.320	92.564	5.451	N.D.
660 minutes (4 theta)	1.231	92.786	5.573	N.D.
					After the reaction	N.D.	94.241	5.400	N.D.

[ Table 7]

Time course of reaction purity in run 5: HPLC area% (230nm)

Time after starting operation	Cyclized precursors	Target molecules	Dimer (cyclized form)	Dimer (straight chain form)
					165 minutes (0.5 theta)	1.567	94.288	3.815	N.D.
330 minutes (theta)	0.650	95.872	3.352	N.D.
					495 minutes (1.5 theta)	0.353	96.429	3.107	N.D.

[ Table 8]

Comparison between simulation and Experimental results (run 3)

In run 3, it was confirmed that the simulation and experimental results were substantially identical.

[ Table 9]

Comparison between simulation and experimental results (runs 4, 5)

The simulation and experimental results were confirmed to be substantially identical in runs 4 and 5.

Calculation formula

Conversion rate of reaction

Simulation: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

The experimental results are as follows: (initial area of cyclized precursor-area of cyclized precursor%)/initial area of cyclized precursor%

Selectivity is

Simulation: target molecule concentration/(target molecule concentration + dimer (straight chain + cyclic) concentration)

The experimental results are as follows: area% of target molecule/(% of target molecule area + dimer (linear + cyclic))

In runs 4 and 5, the area% of dimer decreased due to the presence of side reactions and the area% of the target cyclized form increased compared to those of runs 1-2, which were highly diluted. More specifically, high selectivity was produced with the CSTR and selectivity equal to or higher than the pseudo high dilution of the batch in run 3 could be achieved (table 5).

The use of a CSTR enables the preparation to be carried out using smaller equipment than batch reactions. FIGS. 1 and 2 show a comparison of the size of the equipment used when the reaction is carried out using a CSTR and when a pseudo-high dilution batch reaction is carried out by reverse dropwise addition of the starting materials, when it is assumed that about 2kg of product is produced. In this case, the effective working day to obtain 2kg of target molecule was 1 day in both cases; however, the equipment (particularly the size of the reaction vessel) can be reduced by a factor of about 20. In addition, the use of one or more CSTRs enables continuous operation, and an increase in efficiency of operation (including subsequent processing) can be expected.

[ example 2]Synthesis of cyclosporin A

The scheme is as follows:

synthesis of cyclized precursors

165.2g of open-ring form 1 was synthesized from 198.3g of cyclosporin A according to the non-patent literature (Bernard Riss, Arnaud Grandery, Thorsten Gut, Manuela Seeger-Weibel, Christian Zuercher, Jinpling Li, and Fabrice Gallou. org. Processss. Dev.2014, 18, 1763-1770).

The above ring-opened form 1(10.0g) was dissolved in methanol (300mL), a solution of sodium methoxide in methanol (28% w/w, 5.7g) was added to the resulting solution, and allowed to react. After stirring at room temperature for 14 hours, 1M aqueous sodium hydroxide solution (37mL) was added, and it was further reacted at room temperature for 4 hours. Next, a 1M potassium hydrogen sulfate solution was added to the reaction solution at an ice-cooled temperature for neutralization so that the pH value thereof became about 7. The precipitated solid was filtered, and the filtrate was extracted twice with ethyl acetate (200 mL). The organic layer was then washed with water and concentrated using an evaporator to obtain cyclized precursor 2(8.1g) as a white powder.

Simulation of reaction Rate parameters

The data on the change in concentration of each compound according to temperature/reaction time was collected at 5 ℃, 21 ℃ or-9 ℃ to calculate the reaction rate parameter. Concentration data at 5 ℃ and 21 ℃ were collected using a plug flow reactor. More specifically, a solution of the cyclization precursor (3mM) and DIPEA (16mM) in DMF (solution A) and an 8mM solution of O- (7-azabenzotriazol-1-yl) -1, 1, 3, 3-tetramethyluronium Hexafluorophosphate (HATU) in DMF (solution B) were reacted at 5 ℃ or 21 ℃. The reaction was quenched with methylamine solution. The reaction time is regulated by the flow rates of the solutions A and B. In another aspect, data at-9 ℃ was collected using a batch reactor. More specifically, a reaction solution prepared by dissolving a cyclized precursor (35.0mg, 74% content, 0.02mmol) in DMF (14.3mL) and adding DIPEA (20mL, 5.4 equivalents) and HATU (22.3mg, 2.8 equivalents) thereto was sampled over time. The cyclized precursor is detected as formamide.

The compound concentrations shown in table 10 were calculated from the HPLC data based on HPLC determined values for cyclization precursor and cyclosporin a. The cyclized precursors were detected as formamide forms of the cyclized precursors and their absorption coefficients were considered to be equal. In addition, the absorbances of cyclosporin a and the dimer (cyclized form) were considered to be equal.

[ Table 10]

Data on the variation of the concentration of the Compound (example 2)

Temperature C	Reaction time(sec)	Cyclization precursor (mM)	Cyclosporin A (mM)	Dimer (cyclized form) (mM)
					-9	60	1.12	0.18	0.09
-9	120	0.80	0.32	0.24
					-9	240	0.46	0.52	0.43
5	32	1.03	0.25	0.11
					5	64	0.59	0.43	0.24
5	128	0.33	0.65	0.37
					21	32	0.70	0.52	0.16
21	64	0.42	0.74	0.26
					21	128	0.15	0.96	0.32

From the data obtained, the mechanism of this cyclization reaction is considered as follows. No major effect of the activator on the reaction rate was observed; therefore, it is omitted.

Using the data on the change of concentration with time obtained by the experiment and the temperature data (Table 10), the reaction rate constants k1, k2 and k3 at each temperature (-9 ℃, 5 ℃ and 21 ℃) for each elementary reaction were obtained. The reaction rate constants k1, k2, and k3 were calculated using the following equations:

TM: cyclosporin a; SM: cyclizing the precursor; SM-HATU: a reactive intermediate; dimer: a dimer; c: a concentration (M); k: constant of reaction rate

From the obtained reaction rate constants k1, k2 and k3 at each temperature and the temperatures, the frequency factor (a) and activation energy (E) of each elementary reaction were calculated from the arrhenius curve. The calculation was performed using Microsoft Excel.

As a result, the data shown in table 11 were obtained.

[ Table 11]

Frequency factors (A) and activation energies (E) corresponding to reaction rate constants k1, k2, and k3

	A	E[J/mol]
			k1	4.62E+3[1/sec]	24558
k2	7.56E+4[1/sec]	37403
			k3	8.94E+4[L/mol/sec]	23673

The obtained frequency factor (a) and activation energy (E) of each elementary reaction were substituted into the following arrhenius equation, and by using the above reaction rate equation, reaction rate constants k1, k2, and k3 were calculated in the case where the temperature at which the cyclization reaction was carried out in the CSTR was set to 25 ℃ (table 12).

[ Table 12]

Reaction rate constants k1, k2 and k3 for carrying out cyclization reactions in CSTR

Temperature of	k1	k2	k3
				25℃	2.30E-01	2.12E-02	6.36E+00

Next, using the reaction rate constants k1, k2, and k3 of table 12, the following mass balance equation and reaction rate equation, the reaction conversion, selectivity, and residence time when the cyclized precursor concentration was set to 0.04mol/L and the reaction temperature was set to 25 ℃ were calculated (table 13).

[ Table 13]

Calculated reaction conditions (when the concentration of the cyclized precursor was 0.04mmol/L and the reaction temperature was 25 ℃ C.)

	Conversion rate of reaction	Selectivity	Retention time [ theta, min ]]
				Case 1	0.99	0.94	80
Case 2	0.99	0.97	160

In cases 1 and 2 of Table 13, the residence time that will produce the results that will make the conversion and selectivity of the reaction higher was determined by calculation. Case 2 (run 3) was performed in this example.

Calculation formula

Reaction conversion rate: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

And (3) selectivity: cyclosporin A concentration/(Cyclosporin A + dimer (cyclized form) concentration)

Run 1: batch high dilution

HATU (18.9mg, 2.7 equiv.) was added to a solution of the precursor compound (30.0mg, 74% content, 0.02mmol) and DIPEA (17 μ L, 5.4 equiv.) in DMF (12.3mL) and allowed to react for 1 hour. When the concentration of cyclosporin A in the reaction solution was quantified to calculate the yield, it corresponded to 15.8mg (72% yield).

Run 2: batch pseudo high dilution

To a solution of HATU (311.9mg, 2.7 equivalents) and DMF (10mL) was added a solution of the precursor compound (500.5mg, 74% content, 0.30mmol) and DIPEA (0.3mL, 5.7 equivalents) in DMF (4.7mL) dropwise over a period of 180 minutes. After about 60 minutes, when the concentration of cyclosporin A in the reaction solution was quantified to calculate the yield, it corresponded to 348.3mg (95% yield).

Run 3: CSTR (continuous stirred tank reactor)

The cyclized precursor (1.9009g, 74% content, 1.15mmol) and DIPEA (1.1mL, 5.5 equiv.) were dissolved in DMF (27.4 mL). The substrate concentration was 0.04 mmol/mL. In another aspect, HATU (1.1904g, 2.7 equivalents) was dissolved in DMF (28.5 mL). The reagent concentration was 0.11 mmol/mL. These solutions were added to 12.8mL of DMF solution at 25 ℃ at an input rate of 0.04mL/min, respectively, and the reaction solutions were simultaneously withdrawn at an output rate of 0.08 mL/min. Samples were taken by collecting the reaction solutions 160 minutes (1 θ), 320 minutes (2 θ), 480 minutes (3 θ) and 640 minutes (4 θ) after the start of the operation, and the samples were quenched using dimethylamine solution. All the output solutions and the solution in the reactor were put together after 640 minutes and when the concentrations were quantified to calculate the yield, it corresponded to 1.14g (98% yield).

An increase in the purity of the target molecule was observed at or after the operating time 2 θ in run 3 compared to the pseudo high dilution method in run 2.

Under the conditions of run 3, using a 7 liter reaction kettle (fig. 1, same as example 1), 2kg can be prepared in one day.

Analysis conditions were as follows:

column: BIOshell C18, 2.1mm I.D.x 150mm L, 2.6 μm

Column temperature: 60 deg.C

Flow rate: 0.5mL/min

Gradient (B%): 0 to 40.0min (30 → 100), 40.0 to 40.6min (100), 40.6 to 40.7min (100 → 30), 40.7 to 45.0min (30).

Cyclization precursor:

LCMS(m/z)[M+H]⁺: calculated 1220.8598, found: 1220.8674, respectively; elution time: 11.3 minutes. When the reaction is followed, the cyclized precursor is detected as the formamide present after quenching. Elution time: 11.7 minutes.

And (3) determination: 74% as determined by qNMR and HPLC

a. Sample qNMR:

measurement conditions were as follows: DMSO-d6, 100 ℃, pulse angle: 90 °, digital resolution: 0.15, relaxation time: 60 seconds, no rotation, cumulative number of times: 64 times

1H NMR peak assignments (see FIG. 3): (Me for two Ala, 6H in total), (iPrCH for NH-Val, 1H), identified by COSY and TOCSY.

Internal standard: 3- (trimethylsilyl) -1-propane-1, 1, 2, 2, 3, 3-d 6-sodium sulfonate

The content is as follows: 81.2 percent

b. Results of HPLC measurement using the raw materials used in this example as samples: 91.7 percent

Cyclosporin A:

LCMS：m/z[M+H]⁺: calculated 1202.8492, found: 1202.8589, respectively; elution time: 21.5 minutes.

¹H NMR(CDCl₃): the same information as in the following documents: kessler et al Helvetica Chimicaacta, 1985, 68, 661.

Dimer (cyclization): m/z [ M +2H]²⁺: calculated values: 1203.3509, found: 1203.3517, respectively; elution time: 29.1 minutes

β -Elimination of impurities (also known as β -Elimination of impurities) LCMS M/z [ M + H]⁺: calculated 1202.8492, found: 1202.8587, respectively; elution time: 12.6 minutes

β -Elimination of impurities (cyclized form). LCMS (M/z) [ M + H]⁺: calculated 1184.8386, found: 1184.8448, respectively; elution time: 22.0 minutes

As a result:

[ Table 14]

Results for reaction purity in run 1: HPLC area% (205nm)

Reaction time	Cyclized precursors	Cyclosporin A	Dimer (cyclized form)	β -Elimination of impurities (cyclized form)
					1 hour	0.44	73.12	19.84	6.60

Calculation was performed by limiting the sum of cyclized precursor, cyclosporin a, dimer (cyclized form), and β -eliminating impurity (cyclized form) to 100%.

[ Table 15]

Time course of reaction purity in run 2: HPLC area% (205nm)

Reaction time	Cyclized precursors	Cyclosporin A	Dimer (cyclized form)	β -Elimination of impurities (cyclized form)
					180 minutes after the dropwise addition	0.54	85.10	6.51	7.85
30 minutes after the reaction	0.54	85.28	6.52	7.67

[ Table 16]

Time course of reaction purity in run 3: HPLC area% (205nm)

Time after initiation of reaction	Cyclized precursors	Cyclosporin A	Dimer (cyclized form)	β -Elimination of impurities (cyclized form)
					160 minutes (theta)	0.94	86.16	5.30	7.60
320 minutes (2 theta)	0.59	86.27	5.41	7.74
					480 minutes (3 theta)	0.62	86.61	5.51	7.26
640 minutes (4 theta)	0.29	87.12	4.99	7.60
					After the reaction	0.06	87.14	4.94	7.85

[ Table 17]

Comparison between simulation and Experimental results (run 2)

The simulation and experimental results were confirmed to be substantially identical in run 2.

[ Table 18]

Comparison between simulation and Experimental results (run 3)

In run 3, it was confirmed that the simulation and experimental results were substantially identical.

Calculation formula

Conversion rate of reaction

Simulation: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

The experimental results are as follows: (initial area of cyclized precursor-area of cyclized precursor%)/initial area of cyclized precursor%

Selectivity is

Simulation: cyclosporin A concentration/(Cyclosporin A + dimer (cyclized form) concentration)

The experimental results are as follows: area% of cyclosporin A/(% area of cyclosporin A + area% of dimer (cyclized form))

In run 3 (table 16), the area% of dimer decreased due to the presence of side reactions and the area% of the target cyclized form increased compared to those highly diluted in run 1 (table 14). More specifically, high selectivities were produced with CSTRs and selectivities equal to or higher than the batch pseudo high dilution in run 2 (table 15) could be achieved.

[ example 3]Preparation of cyclosporin A

The reactions were scaled up with reference to the data obtained in example 2.

Pseudo-steady-state was previously prepared by batch pseudo-high dilution and the reaction was carried out using a CSTR, as described herein.

Batch pseudo-high dilution

Insoluble material was observed when the precursor compound (20.6354g, 59% content, 10mmol) was dissolved in DMF (155 mL). Then, the insoluble material was filtered, and after confirming that the precursor compound was not included in the solid, a cyclized precursor solution was prepared by adding DIPEA (10mL, 5.7 equivalents) to the filtrate. To a solution of HATU (10.2745g, 2.7 equivalents) and DMF (330mL) prepared in advance in a 1-liter reaction kettle, the cyclized precursor solution was added dropwise over a period of 195 minutes, and the obtained prepared solution was stored at room temperature for 1 day.

-CSTR

A reaction substrate solution was prepared by: the cyclized precursor (61.902g, 59% content, 30mmol) was mixed with DMF (713mL), the insoluble material was removed, and DIPEA (30mL, 5.7 equiv.) was added. The substrate concentration was 0.04 mmol/mL. A reaction reagent solution was prepared by dissolving HATU (36.145g) in DMF (866 mL). The reagent concentration was 0.11 mmol/mL. The reaction substrate solution and the reaction reagent solution were added to the prepared solution prepared by pseudo-high dilution in batches, respectively, at an input rate of 1.5mL/min, and the reaction solution was simultaneously output at 3.0 mL/min.

Ethyl acetate was mixed into the output line at 6.0mL/min using a T-tube, and then water was mixed at 3.0mL/min using a T-mixer. It was confirmed that the mixed solution underwent interfacial separation in the cartridge, and the upper layer (organic phase) and the lower layer (aqueous phase) were output at 5.5mL/min to 6.5mL/min, respectively. The output flow rate is adjusted so that the liquid separation interface will become constant.

The upper layer (organic phase) was temporarily stored in a surge tank and when the volume reached about 400mL, it was concentrated using an evaporator. The internal temperature during the concentration operation was 40 ℃, and the concentration operation was continued while adding the organic layer with a cycle time of 15 minutes to 20 minutes.

The input to the kettle included adding the entire amount over 550 minutes. The post-treatment (operations of separation and concentration of the reaction solution, similar to those described above) was carried out in parallel with the reaction, and an additional 160 minutes of operation was subsequently required. Finally, 439.86g of a concentrated residue were obtained. As a result of the measurement, the obtained cyclosporin corresponded to 44.2g (92% yield).

Cyclization precursor: the content was determined to be 59% by qNMR and HPLC.

As a result:

[ Table 19]

Time course of reaction purity: HPLC area% (% 205nm)

Calculated by limiting the sum of cyclized precursor, cyclosporin a, dimer (cyclized form) and β -eliminating impurity (cyclized form) to 100%.

Comparison between simulation and experimental results

[ Table 20]

The results of the pseudo high dilution in batches were approximately the same as those of example 2.

[ Table 21]

The simulation and experimental results were confirmed to be substantially consistent with the use of a CSTR. Furthermore, the homology was higher than in example 2.

Calculation formula

Conversion rate of reaction

Simulation: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

The experimental results are as follows: (initial area of cyclized precursor-area of cyclized precursor%)/initial area of cyclized precursor%

Selectivity is

Simulation: cyclosporin A concentration/(Cyclosporin A + dimer (cyclized form) concentration)

The experimental results are as follows: area% of cyclosporin A/(% area of cyclosporin A + area% of dimer (cyclized form))

At this time, by carrying out the reaction in a 1 liter kettle and carrying out the work-up in a continuous manner, 44.2g of cyclosporin a can be prepared in 710 minutes (550 minutes for the reaction and the remaining 160 minutes for the work-up).

[ example 4]]Synthesis of desmopressin

The scheme is as follows:

simulation of reaction Rate parameters

To calculate the reaction rate parameter, data were collected on the change in concentration of each compound as a function of temperature/reaction time for each compound. The cyclization precursor (8.92mM) prepared in a 4: 1 solution of N, N-Dimethylformamide (DMF) and acetic acid (solution A) and a 32mM solution prepared in a 4: 1 solution of DMF and acetic acid (solution B) were reacted at 0 deg.C, 23 deg.C or 40 deg.C. The reaction was quenched with 1% sodium dithionite solution. The reaction was carried out using a plug flow reactor, the reaction time was adjusted by the flow rates of the solutions a and B, and the data shown in table 22 were obtained.

The compound concentration was obtained by assuming that the molar absorbances of the cyclization precursor and the target molecule were the same, and converting the initial concentration of the cyclization precursor (4.46mM, as the reaction mixture solution of the above experiment) to HPLC area%. In addition, the absorbance of the dimer was assumed to be 2 times that of the cyclized precursor, and the concentration was calculated.

[ Table 22]

Data on the variation of the concentration of the Compound (example 4)

Temperature of	Reaction time (sec)	Cyclization precursor (mM)	Target molecule (mM)	Dimer (straight chain + cyclized form) (mM)
					0	2	3.29	1.11	0.03
0	10	2.63	1.67	0.08
					0	40	0.90	3.30	0.13
23	2	3.22	1.22	0.03
					23	10	1.97	2.33	0.08
23	40	0.31	3.93	0.11
					40	2	2.99	1.40	0.04
40	10	1.62	2.66	0.09
					40	40	0.28	3.97	0.11

From the data obtained, the mechanism of this cyclization reaction is considered as follows. The reaction from the cyclized precursor to the iodide intermediate is considered to be rapid; thus, the reaction from the iodide intermediate to the target molecule and to the dimer(s) is considered to be the rate limiting step. Although the concentration of the iodide intermediate cannot be measured, since the iodide intermediate can be returned to the cyclized precursor due to quenching before the analysis, and the above-described reaction rate difference is also taken into consideration, it is no problem to assume that the concentration of the iodide intermediate is equal to the concentration of the cyclized precursor obtained by the analysis. Therefore, each parameter was calculated using the elementary reaction shown below. Since iodine is used in excess relative to the cyclized precursor and no large effect of iodine on the reaction rate is observed, it was omitted in the reaction rate simulation.

Using the concentration change data with time and the temperature data obtained by the experiment (table 22), the frequency factor (a) and the activation energy (E) of each elementary reaction were determined using the following equations:

SM: cyclizing the precursor; TM: a target molecule; dimer: a combination of linear and cyclic dimers; c: a concentration (M); kn: a reaction rate constant; an: a frequency factor; en: activation energy; t: (ii) temperature; r: gas constant

Calculations were performed using an Aspen custom modeler from Aspen technology. As a result, the data shown in table 23 were obtained.

[ Table 23]

Frequency factor (A) and activation energy (E) corresponding to reaction rate constants k1 and k2

	A	E[J/mol]
			k1	36.4614[1/sec]	15004.3
k2	13.3879[L/mol/sec]	4854.01

From the obtained frequency factor (A) and activation energy (E) of each elementary reaction, reaction rate constants k1 and k2 were calculated when the temperature at which the cyclization reaction was carried out in the CSTR was set to 25 ℃ (Table 24).

[ Table 24]

Reaction rate constants k1, k2 at 25 ℃ when cyclization is carried out in a CSTR

Temperature of	k1	k2
			25℃	8.57E-2	1.89

Next, using the reaction rate constants k1 and k2 of table 24, the following mass balance equation and reaction rate equation, the conversion rate, selectivity and residence time of the reaction when the cyclized precursor concentration was set to 004mol/L and the reaction temperature was set to 25 ℃ were calculated (table 25).

[ Table 25]

Calculated reaction conditions (when the concentration of the cyclized precursor was 0.04mol/L and the reaction temperature was 25 ℃ C.)

	Conversion rate of reaction	Selectivity	Retention time [ theta, min ]]
				Case 1	0.994	0.997	30
Case 2	0.997	0.999	60
				Case 3	0.998	0.999	120

In cases 1 to 3 of Table 4, the residence time that will produce the results that will make the conversion and selectivity of the reaction higher was determined by calculation. In the present embodiment, case 2 (run 3) is performed.

Calculation formula

Reaction conversion rate: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

And (3) selectivity: target molecule concentration/(target molecule concentration + dimer (linear + cyclization) concentration)

Run 1: batch high dilution

According to the non-patent literature (Dominic Ormerod, Bart Noten, Matthieu Dorbec, Lars Andersson, Anita Buekenhoudt and Ludwig Goetlen. org. Process Res. Dev.2015, 19, 841. 848.), the cyclized precursor (200.8mg, 86.2% content, 0.143mmol) was dissolved with N, N-Dimethylformamide (DMF) (4.4mL) and acetic acid (1.6mL) and a solution of iodine (4.4 equivalents) in DMF (solution volume: 2.5mL) was added to the mixture at room temperature. The resulting mixture was allowed to react at room temperature for 5 minutes. After quenching the reaction solution with an aqueous solution of sodium dithionite, the solution concentration of the target molecule was quantified, and when the yield was calculated, it corresponded to 121.6mg (80% yield).

Run 2: batch pseudo high dilution

A solution of the cyclized precursor compound (502.0mg, 86.2% content, 0.357mmol), DMF (6mL) and acetic acid (1.5mL) was added dropwise over a period of 60 minutes to a solution of iodine (0.3212g, 3.5 equiv.), DMF (10mL) and acetic acid (2.5mL) at 22 ℃. Approximately 30 minutes after the dropwise addition, when the concentration of the solution of the target molecule was quantified to calculate the yield, it corresponded to 284.4mg (75% yield).

Run 3: CSTR (continuous stirred tank reactor)

The cyclized precursor (1.2040g, 86.2% content, 0.855mmol) was dissolved in DMF (19.2mL) and acetic acid (4.8 mL). The substrate concentration was 0.04 mmol/mL. In another aspect, iodine (0.8497g, 3.348mmol) was dissolved in DMF (20.0mL) and acetic acid (5.0 mL). The reagent concentration was 0.13 mmol/mL. These solutions were added to DMF (9.6 mL)/acetic acid (2.4mL) solutions at an input rate of 0.1mL/min, respectively, at 23 deg.C, and the reaction solutions were simultaneously withdrawn at an output rate of 0.2 mL/min. Samples were taken by collecting the reaction solution at 60 minutes (θ), 120 minutes (2 θ), 180 minutes (3 θ) and 240 minutes (4 θ) after the start of the operation, and these samples were quenched using a 0.05% sodium dithionite solution. When all the output solutions obtained by inputting 97.3% of the starting materials for preparation were put together and the concentration was quantified to calculate the yield, it corresponded to 831.9mg (93% yield).

Theta is elapsed time/mean residence time

Analysis conditions were as follows:

HPLC and LCMS conditions:

column: biochell C18, 2.1mm I.D.x 150mmL, 2.6 μm

Mobile phase: A) water to TFA is 2000 to 1; B) acetonitrile to TFA 2000: 1

Column temperature: 60 deg.C

Flow rate: 0.5mL/min

Gradient (B%): 0 to 20.0min (5 → 45), 20.1 to 23.0min (100), 23.1min and beyond (5).

MS detection mode: ESI (LC/MS): m/z

Target molecule (desmopressin):

LCMS：ESI(m/z)：1069[M+H]⁺(ii) a Elution time: 8.8 minutes

Sample qNMR (trifluoroacetate):

measurement conditions were as follows: DMSO-d6, 75 ℃, pulse angle: 90 °, digital resolution: 0.29Hz, relaxation time: 30 seconds, no rotation, cumulative number of times: 64 times

The CH protons of Gln (1H in total, delta: 3.985-4.021ppm) were used for quantification

Internal standard: 3, 5-bis (trifluoromethyl) benzoic acid

The content is as follows: 77.4 percent

Cyclization precursor:

LCMS：ESI(m/z)：1213[M+H]⁺(ii) a Elution time: 7.6 minutes

Sample qNMR (trifluoroacetate):

measurement conditions were as follows: DMSO-d6, 23 ℃, pulse angle: 90 °, digital resolution: 0.29Hz, relaxation time: 60 seconds, no rotation, cumulative number of times: 8 times (by volume)

α -carbon CH protons (total of 3H, delta: 4.516-4.722ppm) were used for quantification

Internal standard: 3, 5-Dinitrobenzoic acid methyl ester

The content is as follows: 86.2 percent

Dimer (cyclized form-1)

LCMS：ESI(m/z)：1070[M+2H]²⁺(ii) a Elution time: 10.5 minutes

Dimer (cyclized form-2)

LCMS：ESI(m/z)：1070[M+2H]²⁺(ii) a Elution time: 12.2 minutes

By-products:

LCMS：ESI(m/z)：1195[M+H]⁺(ii) a Elution time: 11.6 minutes

As a result:

[ Table 26]

Results of reaction purity in runs 1-3: HPLC area% (210nm)

[ Table 27]

Time course of reaction purity in run 3: HPLC area% (210nm)

[ Table 28]

Comparison between simulation and Experimental results (run 2)

In run 2, some differences were observed between the simulation and experimental results. The reason for this is considered to be that, when the cyclized precursor is dropped on the scale of run 2, the reaction proceeds before it is uniformly dispersed due to the very fast reaction rate characteristic, and it is considered that the simulation result will coincide with the experimental result at the time of the scale-up experiment.

[ Table 29]

Comparison between simulation and Experimental results (run 3)

In run 3, it was confirmed that the simulation and experimental results were substantially identical.

Calculation formula

Conversion rate of reaction

Simulation: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

The experimental results are as follows: (initial area of cyclized precursor-area of cyclized precursor%)/initial area of cyclized precursor%

Selectivity is

Simulation: target molecule concentration/(target molecule concentration + dimer (straight chain + cyclic) concentration)

The experimental results are as follows: area% of target molecule, + area% of by-product/(% of target molecule, + area% of by-product + dimer (linear + cyclic))

(since the by-product is formed by further iodination of the target molecule, it is calculated together with the target molecule).

[ example 5]Synthesis of cyclosporin A derivatives

The scheme is as follows:

synthesis of cyclic precursor 5

Cyclized precursor 5 was synthesized from the above-mentioned compound 2 (cyclized precursor 2 in example 2) in 3 steps.

Preparation of Compound 3

Compound 2(7.1g) above was dissolved in N, N-dimethylformamide (71mL), and N, N-diisopropylethylamine (3mL) and chloroacetyl chloride (1.85mL) were added on ice. After stirring at room temperature for 2 hours, a 5% potassium hydrogensulfate solution (100mL) and ethyl acetate (100mL) were added to the reaction solution, followed by liquid separation. The organic layer was concentrated using an evaporator, and then purified by silica gel chromatography (ethyl acetate/heptane/methanol) to obtain compound 3(5.0g) as a yellow oily liquid.

LCMS：ESI(m/z)：1297[M+H]⁺

Preparation of Compound 4

Compound 3(5.0g) obtained in the previous step was dissolved in N, N-dimethylformamide (50mL), and S-trityl-L-cysteine methyl ester hydrochloride (4.0g), HOAt (1.6g), and EDC hydrochloride (2.6g) were added. After stirring at room temperature for 2 hours, a saturated aqueous solution of sodium hydrogencarbonate (100mL) and ethyl acetate (100mL) were added to the reaction solution, and liquid separation was performed. The organic layer was concentrated using an evaporator, and then purified by silica gel chromatography (ethyl acetate/heptane/methanol) to obtain compound 4(4.7g) as a yellow oily liquid.

LCMS：ESI(m/z)：1656[M+H]⁺

Preparation of cyclized precursor 5

Compound 4(4.7g) obtained in the previous step was dissolved in dichloromethane (47mL), and triisopropylsilane (3mL) and trifluoroacetic acid (5.5mL) were added on ice. After stirring at room temperature for 1 hour, the reaction solution was concentrated using an evaporator, and then the residue was purified by silica gel chromatography (ethyl acetate/heptane/methanol/trifluoroacetic acid). Further purification by reverse phase chromatography (water/acetonitrile/trifluoroacetic acid) gave the cyclized precursor 5(2.1g) as a white powder.

Simulation of reaction Rate parameters

Concentration variation data for each compound was collected based on the temperature/reaction time for each compound to calculate the reaction rate parameter. To the cyclized precursor (4.63mM) prepared in N, N-Dimethylformamide (DMF) solution was added N, N-Diisopropylethylamine (DIPEA) (9.9 to 10.9 equivalents), and it was reacted at 3 deg.C, 24 deg.C or 46 deg.C. The reaction was quenched with 0.5% TFA in acetonitrile-water (1: 1). The reaction was followed by HPLC over time and the data shown in table 30 was obtained. The concentration of each compound was obtained by converting the concentration of the cyclized precursor (4.63mM) to HPLC area%. The absorption coefficient of the dimer is assumed to be 2 times that of the cyclized precursor, and the absorption coefficient of the trimer is assumed to be 3 times that of the cyclized precursor, to calculate the concentration.

[ Table 30]

Data on the variation of the concentration of the Compound (example 5)

From the data obtained, the mechanism of this cyclization reaction is considered as follows. The reaction from the cyclized precursor and DIPEA to the intermediate is sufficiently fast; thus, the rate of production of the target molecule and dimer and trimer(s) from the intermediate is considered to be the rate limiting step. Although the concentration of the intermediate could not be measured, it was not problematic to calculate each parameter by: it is assumed that the intermediate concentration is equal to the starting material concentration obtained by the analysis (since it is likely that the intermediate can be returned to the cyclization precursor by quenching before the analysis), and the above-described reaction rate difference is also taken into account. In addition, since DIPEA is used in excess relative to the cyclization precursor and its influence on the reaction rate is small, it is omitted.

Using the concentration change data with time and the temperature data obtained by the experiment (table 30), the frequency factor (a) and the activation energy (E) of each elementary reaction were determined using the following equations:

SM: cyclizing the precursor; IM: an intermediate; TM: a target molecule; dimer: the sum of dimers and trimers; c: a concentration (M); kn: a reaction rate constant; an: a frequency factor; en: activation energy; t: (ii) temperature; r: gas constant

Calculations were performed using an Aspen custom modeler from Aspen technology. As a result, the data shown in table 31 were obtained.

[ Table 31]

Frequency factor (A) and activation energy (E) corresponding to reaction rate constants k1 and k2

	A	E[kJ/mol]
			k1	NA	NA
k2	82906.5	28071.8
			K3	63.1182	108.973

From the obtained frequency factor (A) and activation energy (E) of each elementary reaction, reaction rate constants k2 and k3 were calculated when the temperature at which the cyclization reaction was carried out in the CSTR was set to 25 ℃ (Table 32).

[ Table 32]

Reaction rate constant at 25 ℃ when cyclization is carried out in a CSTR

Temperature of	k1	k2	k3
				25℃	NA	1.00	60.4

Next, using the reaction rate constants k2 and k3 of table 32, the following mass balance equation and reaction rate equation, the reaction conversion, selectivity and residence time when the cyclized precursor concentration was set to 0.027mol/L and the reaction temperature was set to 25 ℃ were calculated (table 33).

[ Table 33]

Calculated reaction conditions (when the cyclized precursor concentration was 0.027mol/L, the reaction temperature was 25 ℃ C.)

In cases 1 to 3 of Table 33, the residence time which will produce the results that will make the conversion and selectivity of the reaction higher was determined by calculation. Case 3 is performed in this embodiment.

Calculation formula

Reaction conversion rate: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

And (3) selectivity: target molecule concentration/(target molecule concentration + dimer (dimer + trimer) concentration)

Run 1: batch high dilution

The cyclized precursor (49.7mg, 77.3% content, 0.0272mmol) was dissolved in DMF (8.8mL) and DIPEA (0.28mmol, 10 equiv.) was added to the mixture at room temperature. The resulting mixture was allowed to react at room temperature for 1 hour. A portion of the reaction solution was quenched with 0.5% trifluoroacetic acid in acetonitrile-water (1: 1) solution and it corresponded to 30.8mg (82% yield) when quantifying the concentration of the target molecule in solution to calculate the yield.

Run 2: batch pseudo high dilution

To a solution of DIPEA (1.72mmol, 10 equiv.) in DMF (7.5mL) at 22 ℃ a solution of the cyclized precursor compound (300.9mg, 77.3% content, 0.164mmol) in DMF (4.6mL) was added dropwise over a period of 3 hours. Approximately 1 hour after the dropwise addition, when the concentration of the solution of the target molecule was quantified to calculate the yield, it corresponded to 187.3mg (83% yield).

Run 3: CSTR (continuous stirred tank reactor)

The cyclized precursor (1.3671g, 77.3% content, 0.747mmol) was dissolved in DMF (27.3 mL). The substrate concentration was 0.027 mmol/mL. In another aspect, DIPEA (773mmol, 10 equiv.) was dissolved in DMF (27.3 mL). The concentration of the reagent was 0.283 mmol/mL. These solutions were added to DMF (13.2mL) at an input rate of 0.04mL/min, respectively, at 23 ℃ and the reaction solutions were simultaneously withdrawn at an output rate of 0.08 mL/min. Samples were taken by collecting the reaction solution 165 minutes (theta), 330 minutes (2 theta), 495 minutes (3 theta) and 660 minutes (4 theta) after the start of the operation, and quenched with 0.5% trifluoroacetic acid in acetonitrile-water (1: 1) solution. When all output solutions were put together and the concentrations quantified to calculate the yield, it corresponded to 793.9mg (82% yield). The yield was calculated based on the amount of the raw material solution used for input (94.5% of the total amount).

Theta is elapsed time/mean residence time

Analysis conditions were as follows:

HPLC and LCMS conditions:

column: kinetex Biphenyl, 2.1mm I.D.x.150 mm, 2.6 μm

Mobile phase: A) water to TFA is 2000 to 1; B) acetonitrile to TFA 2000: 1

Column temperature: 60 deg.C

Flow rate: 0.5mL/min

Gradient (B%): 0 to 30min (10 → 100), 30.0 to 30.6min (100), 30.7min and beyond (10).

MS detection mode: ESI (LC/MS): m/z

Target molecule (cyclosporin a derivative):

LCMS：ESI(m/z)：1378[M+H]⁺(ii) a Elution time: 17.5 minutes

The yield was quantified by HPLC assay using the following samples.

Sample qNMR:

measurement conditions were as follows: DMSO-d6, 147 ℃, pulse angle: 90 °, digital resolution: 0.25Hz, relaxation time: 60 seconds, no rotation, cumulative number of times: 32 times (twice)

The β -carbon CH protons of MeVal (total of 1H) were used for quantification

Internal standard: maleic acid

The content is as follows: 94.4 percent

Cyclization precursor:

LCMS：ESI(m/z)：1414[M+H]⁺elution time: 13.9 minutes

The content is as follows: 77.3% as determined by qNMR and HPLC measurements

a. Sample qNMR:

measurement conditions were as follows: DMSO-d6, 90 ℃, pulse angle: 90 °, digital resolution: 0.25Hz, relaxation time: 60 seconds, no rotation, cumulative number of times: 32 times (twice)

The β -carbon CH protons of MeVal (total of 1H) were used for quantification

Internal standard: 3, 5-bis (trifluoromethyl) benzoic acid

The content is as follows: 91.8 percent

b. Results of HPLC measurement using the raw materials used in this example as samples: 84.2 percent of

Dimer

LCMS：ESI(m/z)：1378[M+2H]²⁺Elution time: 21.8 minutes

Trimer

LCMS：ESI(m/z)：1378[M+3H]³⁺Elution time: 24.1 minutes

As a result:

[ Table 34]

Results of reaction purity in runs 1-3: HPLC area% (210nm)

[ Table 35]

Time course of reaction purity in run 3: HPLC area% (210nm)

[ Table 36]

Comparison between simulation and Experimental results (run 2)

In run 2, some differences were observed between the simulation and experimental results. The reason for this is considered to be that, when the cyclized precursor is dropped on the scale of run 2, the reaction proceeds before it is uniformly dispersed due to the very fast reaction rate characteristic, and it is considered that the simulation result will coincide with the experimental result at the time of the scale-up experiment.

[ Table 37]

Comparison between simulation and Experimental results (run 3)

In run 3, it was confirmed that the simulation and experimental results were substantially identical.

Calculation formula

Conversion rate of reaction

Simulation: (initial concentration of cyclization precursor-concentration of cyclization precursor)/initial concentration of cyclization precursor

The experimental results are as follows: (initial area of cyclized precursor-area of cyclized precursor%)/initial area of cyclized precursor%

Selectivity is

Simulation: target molecule concentration/(target molecule concentration + (dimer + trimer) concentration)

The experimental results are as follows: (area of target molecule%)/(area of target molecule% + (dimer + trimer)%)

INDUSTRIAL APPLICABILITY

The present invention provides a novel process for the preparation of cyclic organic compounds, said process comprising carrying out a cyclization reaction using one or more CSTRs, and the like. The invention provided by the invention can be used for continuously preparing the cyclic organic compound in one or more smaller reaction kettles and with less impurities.