KR100277206B1 - Resin containing heteroelement-containing aliphatic linkage useful for combinatorial chemical synthesis - Google Patents

Abstract

The present invention relates to a resin containing a heteroelement-containing aliphatic linker, and more particularly, to tracking the progress of a synthesis reaction on a solid support for combinatorial chemistry or to quantifying a synthetic compound. The present invention relates to a novel resin containing a complex element-containing aliphatic linker represented by the following Chemical Formula 1 which is useful for efficiently tracking by using X-ray Photoelectron Spectroscopy (XPS) elemental analysis in performing qualitative analysis. .

In Formula 1 above: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5.

Description

Resin containing heteroelement-containing aliphatic linkage useful for combinatorial chemical synthesis

The present invention relates to a resin containing a heteroelement-containing aliphatic linker, and more particularly, to tracking the progress of a synthesis reaction on a solid support for combinatorial chemistry or to quantifying a synthetic compound. The present invention relates to a novel resin containing a complex element-containing aliphatic linker represented by the following Chemical Formula 1 which is useful for efficiently tracking by using X-ray Photoelectron Spectroscopy (XPS) elemental analysis in performing qualitative analysis. .

Formula 1

In Formula 1 above: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5.

Combinatorial Chemical Synthesis (hereinafter referred to as "CCS") is a new field of research in the development of new materials and materials. CCS is more diverse than traditional classical organic synthesis in one reaction. It is a new concept of chemical synthesis that synthesizes a large number of compounds at the same time. Thus, the introduction of CCS makes it easier to search for new compounds of lead compounds and to optimize their structure and function. In addition, the combination chemical synthesis (CCS) is mostly carried out on a solid support, it is possible to automate the continuous multi-step reaction and reaction process, and the separation and purification process of the product is very simple to enable high throughput screening (HTS) Has the advantage.

As described above, the combination chemical synthesis (CCS) is a new synthesis method that overcomes the inefficiency and inefficiency of the existing synthesis technology, but it was not easily applied to the field of organic synthesis. This is because it was not easy to track the reaction progress according to the reaction time.

In general, since the combinatorial chemical synthesis (CCS) is performed on a solid support, the progress of the reaction is mainly based on solid-nuclear magnetic resonance spectroscopy (Solid-NMR), mass spectrometry (Mass), and infrared spectroscopy (IR). Solid-NMR spectroscopy has the advantage of revealing the overall structure of the compound, but until now it is difficult to obtain clear analytical data to analyze compounds on a solid support broadly. There is a limitation in confirming the structure of the compound having hydrogen and carbon in the same spectral region and quantitatively analyzing the progress of the reaction. Mass spectrometry is effective for the use of small amounts of samples and qualitative analysis of the product, but there are limitations in quantitative analysis and the progress of the reaction. Qualitative analysis is also difficult. In the case of infrared analysis (IR), since qualitative and general quantitative analysis is possible simply, it is most widely used, but there is a disadvantage that almost no quantitative analysis is possible when a plurality of identical functional groups exist in a molecule.

While the above assays can be used in part to analyze compounds on solid supports, it is not possible to quantitatively analyze the progress of the entire chemical reaction on solid supports. Thus, although the chemical reaction on the solid support is performed in the same manner as the liquid chemical reaction, it is difficult to efficiently select the end point of the reaction, and thus has a problem in that the reaction is terminated after a long time.

The present inventors have repeatedly studied to introduce a combinatorial chemistry approach and a high-efficiency mass assay (HTS) methodology that can search a large amount of organic synthesis in the field of organic synthesis, and as a result XPS in the progress of chemical reaction and compound analysis on a solid support (X-ray Photoelectron Spectroscopy) was found to be effective, and in order to effectively utilize this XPS analysis, a new structure of halogenated Wang resin, which is effective for XPS analysis and has excellent chemical stability and reactivity, has already been applied for patent [ Patent Application No. 98-33798].

On the other hand, the XPS assay introduced by the present invention to track the progress of the chemical reaction has been mainly applied in the field of polymer synthesis. There is a problem that it is difficult to use the amount of the element of the solid support as the amount of the standard element because the relative difference in the amount of the elements in the portion is too large (per 1-2 millimoles / g of resin).

In addition, since most of the biologically active substances contain hetero elements, in the present invention, the relative changes of the hetero elements with the progress of the chemical reaction are introduced by introducing a standard hetero element into Merrifield resin, which is a kind of solid support. Accurately tracked and increased the useful value of XPS elemental analysis.

Accordingly, an object of the present invention is to provide a resin containing a heteroelement-containing aliphatic linker useful for combinatorial chemical synthesis and a method for preparing the same. In addition, the present invention is another object to provide a method for quantitative and qualitative analysis of the resin (resin) containing a hetero-element-containing aliphatic linker (Linker) and the product produced during the combinatorial chemical synthesis process by XPS analysis.

Figure 1a shows the results of XPS elemental analysis of the 4-chlorobenzoic acid ester ethyl thio ether resin (Formula 5a),

Figure 1b shows the results of the XPS element analysis for the 4-bromobenzoic acid ester ethyl thio ether resin (Formula 5b) synthesis reaction,

Figure 1c shows the results of XPS element analysis for the 2-iodobenzoic acid ester ethyl thio ether resin (Formula 5c) synthesis reaction,

Figure 2a shows the XPS elemental analysis results for the synthesis of Boc-Gly-ethyl thio ether resin (Formula 6a),

Figure 2b shows the results of XPS elemental analysis for Gly-ethyl thio ether resin (Formula 7a) synthesis reaction,

Figure 2c shows the results of XPS elemental analysis for the synthesis of Boc-Gly-2,2,3,3-tetrafluorobutyl ether resin (Formula 6b).

The present invention is characterized by a resin containing a heteroelement-containing aliphatic linker represented by the following Chemical Formula 1 useful for combinatorial chemical synthesis.

Formula 1

In Formula 1 above: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5.

Referring to the present invention in more detail as follows.

According to the present invention, a method for preparing a resin containing a heteroelement-containing aliphatic linker represented by Formula 1 includes an aliphatic compound containing a heteroelement represented by the following Formula 2, as shown in Scheme 1 below. It consists of a process of preparing a resin containing a hetero-element-containing aliphatic linker represented by the formula (1) by reacting the linker (Merrifield resin) represented by the formula (3) .

In Scheme 1: ⓟ, A, B, X, Y and n are as defined above, respectively.

Introduction reaction of Merrifield resin according to Scheme 1 is carried out in the presence of a base, using a conventional organic solvent, such as dichloromethane as a reaction solvent, the reaction is carried out smoothly under room temperature conditions and heated if necessary It is also possible. In this case, the base may be an inorganic base selected from alkoxy sodium and potassium carbonate having 1 to 4 carbon atoms such as sodium hydride, methoxy sodium, ethoxy sodium or isopropoxy sodium, or alkyl such as triethylamine and diisopropylethylamine. Organic bases selected from amines are used. As a result of the reaction, no residual Merrifield resin was detected because no chlorine ions were detected in the XPS elemental analysis. In addition, the aliphatic linker represented by the formula (2) used in the present invention is a known compound, and may be represented by aliphatic alcohol, aliphatic thiol, and aliphatic amines according to a combination of A and B.

On the other hand, the present invention is produced in a combination chemical synthesis (CCS) process using a new resin (resin) containing a hetero element-containing aliphatic linker (Linker) represented by the formula (1) prepared by Scheme 1 as a solid support It includes the method of accurately tracking the progress of the reaction through the relative quantitative analysis of the compound by X-ray Photoelectron Spectroscopy (XPS) elemental analysis. In this case, the combination chemical synthesis process includes all of the conventional organic synthesis process, peptide synthesis process and carbohydrate synthesis process.

Scheme 2 below shows an example of a conventional organic synthesis process for introducing various types of halobenzoic acid such as chloro, bromo and iodo represented by the formula (4) on the novel resin represented by the formula (1).

In Scheme 2: ⓟ, A, B, X, Y and n are as defined above, respectively; Z is a halogen atom or a hetero atom, which may be substituted at the o-, m- and p-positions; m is the number of substituents Z and is an integer of 0 or 1-5.

According to Scheme 2, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (hereinafter referred to as "EDC") and a catalytic amount of 4-dimethylaminopyridine (hereinafter referred to as "DMAP") ) In the presence of the compound (1) containing a hetero element-containing aliphatic linker (Linker) and the halobenzoic acid is reacted at room temperature. At this time, halobenzoic acid and EDC, it is preferable to use 3 to 5 equivalent times with respect to the resin (resin) containing a hetero element-containing aliphatic linker (Linker) represented by the formula (1), respectively.

In order to track the production of the product according to Scheme 2, a reaction sample was taken and XPS elemental analysis was performed. In the case of Scheme 2, XPS elemental analysis shows that the ratio of the hetero atom Z to the substituent (A or B) on the solid support is 1: 1, or the ratio of the hetero atom Z to the number of substituents (X and Y) is A constant integer ratio is the end point of the reaction. More detailed reaction conditions and relative element analysis results of XPS are described in detail in Example 2 below.

As another example, the following Scheme 3 is an embodiment of applying a resin containing a complex element-containing aliphatic linker represented by the formula (1) in the synthesis of peptide, repeat introduction of glycine (Glycine) The reaction is shown.

In Scheme 3: ⓟ, A, B, X, Y and n are as defined above, respectively; R ¹ represents a commercial amino acid side chain such as hydrogen, methyl, ethyl, isopropyl and the like.

According to Scheme 3, the resin represented by Chemical Formula 1 in the presence of EDC and DMAP and glycine protected by t-butoxycarbonyl group (hereinafter referred to as “Boc-Gly”) are reacted at room temperature for 12 hours. To obtain the compound represented by Chemical Formula 6. The compound represented by Chemical Formula 6 was reacted with a dichloromethane solution of 5-50% trifluoroacetic acid at 0 ° C. to remove t-butoxycarbonyl (hereinafter referred to as “Boc”), and then 50% triethyl The reaction is carried out in a 1,4-dioxane solution of amine to obtain an amino compound represented by the formula (7). Thereafter, various amino acids protected with Boc are reacted with the compound represented by Formula 7 in the presence of EDC and HOBt to obtain a dipeptide compound represented by Formula 8.

In addition, the degree of generation of peptides according to Scheme 3 was traced through XPS elemental analysis. As the number of amino acids increased, the ratio of nitrogen to standard heteroelements increased. When thioethanol was used as the aliphatic linkage, it was confirmed that the ratio of sulfur to nitrogen increased quantitatively with the number of glycine. More detailed reaction conditions and relative element analysis results of XPS are described in detail in Example 3 below.

As described above, complex compounds, peptide synthesis, and the like, are prepared by using a novel resin, which is newly introduced with a complex element-containing aliphatic linker effective in elemental analysis of XPS and excellent in chemical stability and reactivity. When various organic synthesis is performed, the production of the produced compound and the progress of the reaction can be quantitatively and qualitatively confirmed through elemental analysis of XPS.

Such a present invention will be described in more detail based on the following examples, but the present invention is not limited thereto.

Example 1 Preparation of Resin Containing Heteroelement-containing Aliphatic Linker

i) Preparation of Ethanol Thio Ether Resin (Formula 1a)

Triethylamine (4.45 g, 44.0 mmol) with 2-mercapto-1-ethanol (3.13 g, 40.0 mmol) and chloromethyl resin (Merrifield resin, 2 mmol chloromethyl reactor, Formula 3; 4.00 g, 8.0 mmol) Was added to a solution of dichloromethane (100 mL), followed by stirring at room temperature for 48 hours. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol, dichloromethane, dried and analyzed by IR and XPS.

IR; 3327, 1600, 1492, 1451 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p

Ii) Preparation of Propanol Thio Ether Resin (Formula 1b)

Triethylamine (4.45 g, 44.0 mmol) was added 3-mercapto-1-propanol (3.87 g, 40.0 mmol) and chloromethyl resin (Merrifield resin, 2 mmol chloromethyl site, Formula 3; 4.00 g, 8.0 mmol). Was added to a solution of dichloromethane (100 mL), followed by stirring at room temperature for 48 hours. After the reaction was completed, the reaction mixture was filtered, washed repeatedly with water, methanol and dichloromethane, dried and analyzed by IR and XPS.

IR; 3326, 1599, 1492, 1451 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p

Iii) Preparation of 2,2,3,3-tetrafluorobutanol resin (Formula 1c)

Sodium hydride (0.95 g, 3.96 mmol) was added to a solution of 2,2,3,3-tetrafluorobutanediol (3.21 g, 19.8 mmol) in N, N-dimethylformamide (100 mL) and stirred at room temperature for 30 minutes. Then, chloromethyl resin (Merrifield resin, 2.0 mmol chloromethyl site, Chemical Formula; 3.00 g, 6.0 mmol) was added thereto, followed by stirring at 90 ° C. for 48 hours. The reaction end point was when chlorine was not detected. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol, dichloromethane, dried and analyzed by IR and XPS.

IR; 3441, 1602, 1492, 1449 cm ^-1

XPS; 686 eV, F 1s

Example 2 Application of XPS Relative Element Assay to Organic Synthesis Using Ethanol Thioether Resin

i) Reaction of 4-chlorobenzoic acid into ethanol thio ether resin (Formula 5a)

Dichloromethane (15 mL) was synthesized in ethanol thio ether resin (0.20 g, 0.4 mmol; Formula 1a), 4-chlorobenzoic acid (0.19 g, 1.2 mmol), EDC (0.25 g, 1.3) mmol) and catalytic amount DMAP (12.2 mg, 1 × 10 ⁻¹ mmol) were added, followed by stirring at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol, THF, dichloromethane solvent, dried and subjected to IR and XPS elemental analysis.

IR; 1720, 1592, 1490, 1449 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p; 199 eV, Cl 2p

The XPS elemental quantitative analysis results are shown in Table 1A and FIG. 1A, and it is considered that the reaction was terminated as the result of quantitative analysis of S: Cl was 48.3: 51.7.

Response time (hours) S: Area ratio of Cl (%) 12 48.3: 51.7

Ii) Reaction of 4-bromobenzoic acid to ethanol thio ether resin (Formula 5b)

In dichloromethane (15 mL), ethanol thio ether resin (0.20 g, 0.4 mmol; Formula 1a), 4-bromobenzoic acid (0.24 g, 1.2 mmol), EDC (0.25 g, 1.3 mmol) and catalytic amount DMAP (12.2 mg, 1 × 10 ⁻¹ mmol) were added, followed by stirring at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol, THF, dichloromethane solvent, dried and subjected to IR and XPS elemental analysis.

IR; 1719, 1588, 1490, 1450 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p; 190, 184 and 70 eV, Br 3p and 3d

XPS elemental quantitative analysis results are shown in the following Table 1b and Figure 1b, it can be seen that the reaction is terminated as the relative element quantitative analysis of S: Br is 50.7: 49.4.

Response time (hours) S: Area ratio of Br (%) 12 50.7: 49.4

Iii) Reaction of 2-iodobenzoic acid into ethanol thioether resin (Formula 5c)

In dichloromethane (15 ml), ethanol thio ether resin (0.20 g, 0.4 mmol; Formula 1a), 2-iodobenzoic acid (0.29 g, 1.2 mmol), EDC (0.25 g, 1.3 mmol) and catalytic amount DMAP (12.2 mg, 1 × 10 ⁻¹ mmol) were added, followed by stirring at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol, THF, dichloromethane solvent, dried and subjected to IR and XPS elemental analysis.

IR; 1726, 1599, 1582, 1490, 1450 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p; 619 eV, I 3d

XPS elemental quantitative analysis results are shown in the following Table 1c and Figure 1c, the relative element quantitative analysis of S: I is 52.9: 47.1, it can be seen that the reaction progress rate is 95%.

Response time (hours) S: Area ratio of I (%) 12 52.9: 47.9

Example 3 Application of XPS Relative Element Assay to Peptide Synthesis Using Ethanol Thioether Resin

i) Preparation of Boc-Gly-Ethanol Thio Ether Resin (Formula 6a) and XPS Elemental Determination

In dichloromethane (50 mL), ethanol thio ether resin (1.00 g, 2.0 mmol; Formula 1a), Boc-Gly (0.77 g, 4.4 mmol), EDC (0.84 g, 4.4 mmol) synthesized in Example 1 i) ) And catalytic amount DMAP (61.1 mg, 5 × 10 ⁻¹ mmol) were added, followed by stirring at room temperature for 12 hours. After completion of the reaction, the reaction mixture was filtered, washed repeatedly with water, methanol and dichloromethane, dried and subjected to IR and XPS elemental analysis.

IR; 1754, 1712, 1601, 1493, 1450 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p; 402 eV, N 1s

XPS elemental quantitative analysis results are shown in the following Table 2a and FIG. 2a.

Response time (hours) S: area ratio of N (%) 12 49.0: 51.0

Ii) Preparation of Gly-Ethanol Thioether Resin (Formula 7a) and Quantification of XPS Elements

The resin represented by Chemical Formula 6a synthesized in i) of Example 3 was added to dichloromethane (10 ml) in a dichloromethane solution of 50% trifluoroacetic acid, which is a known method, at 0 ° C. for 2 hours to remove the Boc group. , Neutralized with a 1,4-dioxane solution of 50% triethylamine to prepare Gly-ethanol thio ether resin, followed by drying, and then subjected to IR and XPS elemental analysis.

IR; 1740, 1600, 1491, 1450 cm ^-1

XPS; 229 and 165 eV, S 2s and 2p; 402 eV, N 1s

XPS elemental quantitative analysis results are shown in the following Table 2b and FIG. 2b.

Response time (hours) S: area ratio of N (%) 2 48.1: 51.9

Iii) Preparation of Boc-Gly-2,2,3,3-tetrafluorobutanol resin (Formula 6b) and determination of XPS elements

Tetrafluorobutanol resin (0.20 g, 4 × 10 ⁻¹ mmol; Formula 1c), Boc-Gly (0.23 g, 1.32 mmol), EDC (10 mL) synthesized in Example 1) 0.19 g, 1.32 mmol) and HOBt (0.18 g, 1.32 mmol) were added, followed by stirring at room temperature for 12 hours. After the reaction was completed, the reaction product was filtered, washed repeatedly with water, methanol and dichloromethane, dried, and analyzed by IR and XPS elements.

IR; 1722, 1603, 1492, 1450 cm ^-1

XPS; 686 eV, F 1s and 402 eV, N 1s

XPS elemental quantitative analysis results are shown in Table 2c and FIG. 2c.

Response time (hours) F: area ratio of N (%) 12 81.2: 18.8

As is clear from the above examples, in the present invention, combinatorial chemical synthesis is carried out using a novel resin having a linker containing a hetero element equal to the amount of elements in the reactor portion as a solid support. The progress of the reaction can be quantitatively and qualitatively analyzed by XPS elemental quantitative analysis.

Accordingly, the present invention facilitates the application of combinatorial chemical compound synthesis to the search for new compounds with new structures and to optimize their structure and function.

Claims (5)

Resin containing a hetero element-containing aliphatic linker represented by the following formula (1), characterized in that it is useful for combinatorial chemical synthesis. Formula 1 In Formula 1 above: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5. Next, the heteroelement-containing aliphatic compound represented by the following Chemical Formula 1 is prepared by reacting an aliphatic linker containing a linker represented by the following Chemical Formula 2 with Merrifield resin represented by the following Chemical Formula 3. Method for producing a resin (resin) containing a linkage. Formula 1 In the formula: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5. The resin of claim 2, wherein the introduction of the Merrifield resin is carried out in the presence of a base selected from sodium hydride, alkoxy sodium and alkylamine. Way. In the method for analyzing a product by combinatorial chemical synthesis on a solid support, a combinatorial chemical synthesis process using a resin containing a heteroelement-containing aliphatic linkage represented by Formula 1 as a solid support is carried out using XPS (X-ray). Photoelectron Spectroscopy) A method for analyzing a product of a combinatorial chemical synthesis process characterized by quantitative and qualitative analysis by elemental analysis. Formula 1 In Formula 1 above: ⓟ represents polystyrene-divinylbenzene; A and B represent the same or different heteroatoms selected from oxygen, sulfur and nitrogen atoms; X and Y are the same as or different from each other and represent a hydrogen atom or a halogen atom; n is an integer of 1-5. The method of claim 4, wherein the combinatorial chemical synthesis process is an organic synthesis process, a peptide synthesis process or a carbohydrate synthesis process.