DE1804021A1 - Peptide synthesis method - Google Patents

Description

180 * 021

Dn H. Mo--;

Patents /.- 'ill ·; ".

2 HAMBU tx G:

Sivscstr. β, Fo: - ^: - h t-'ϊt -

bi; aao3 October 15, 1968

M.A. 530/68

Client's characters: SBR -

BLACK BIORESEARCH

Division of Becton, Dickinson and Company

Mountain View Avenue

Orangeburg, New York / V.St.A.

Priority: October 26, 1967 / V.St.A. Registration no. 678.187

Peptide synthesis method

The invention relates to a solid phase synthesis process for the preparation of peptides in which the peptide linked to a pesticide carrier via an ester bond constructed and chemically separated from the solid support after construction, and relates to a new and improved one Method for separating the peptide from the solid

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carrier, in which the resin can be reused at the same time. can be reactivated during peptide synthesis.

A known solid phase synthesis method for the preparation of peptides is described by R.B, Merrifield 19β2 ( in Federation Proceedings 21, 412), and the general concept on which this method is based was detailed in I9635 by R.B, Merrifield in J. Am. Chem. Soc. 85, 2149. In short, there is this method consists of building up the peptide chain while one end of the chain is on a carrier is firmly attached, and then separates the synthesized peptide chain from the solid support *

In detail, this process consists in that the first amino acid of the desired peptide is linked to a contaminant carrier via a 0-. "Terminal Car", riyl group. The solid support used must be insoluble in all reagents used in the process, have a large effective surface area and contain a group or be capable of being substituted with a group by which the peptide chain The solid carrier generally used is a 2% crosslinked styrene-divinylbenzene copolymer, which is then attached to the aromatic rings with chloromethyl groups

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is substituted as the reactive group to which the peptide group can be anchored. Then the first amino acid for the desired chain linked to the carrier in such a way that the triethylammonium salt of N-acylamino acid reacted with the chloromethylated polymer support and an ester linkage between the terminal C-atom of the amino acid and the carrier produces.

Then the peptide chain is synthesized in such a way that the protecting Boc group is removed with hydrochloric acid in acetic acid and then the resulting Hydrochloride neutralized with an excess of triethylamine. The unprotected amino acid linked to the solid support then becomes the next amino acid of the desired one Chain coupled, whereby one is protected with a Boc group with the adhering to the carrier Amino acid in the presence of a suitable coupling agent such as dicyclohexylcarbodiimide, brings in contact. The other amino acids of the desired chain are then in the same way, i.e. by removal the Boc group and subsequent domes.

After the desired chain has been synthesized, the peptide is separated from the plague carrier, and this takes place according to the known method according to Merrifield in the manner that a stream of hydrogen bromide through a suspension of the adhered to the support peptide in Trifluoroacetic acid passes through it. Then you can

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treat the set peptide further, e.g. remove the protective group on the last acid of the chain, Hydrogenate to obtain arginine residues from nitroarginine, and the resulting product can be used to obtain the desired Purify peptides »

Although this is a well-known solid synthesis method for production of peptides very useful, but the overall effectiveness of this method leaves much to be desired, since the Separation of the peptide from the pesticide carrier is remarkable Brings difficulties. Under the conditions used chain degradation takes place and this reduces the yield of the desired product. In addition, in in the separation stage a not inconsiderable amount of the desired product on the solid carrier, whereby the Overall yield further reduced. In addition, the chemicals used for this separation are both toxic and toxic difficult to handle, which additionally affects the known method.

The invention is based on the object of eliminating these disadvantages. This object is achieved by means of a solid phase synthesis process the type specified for the production of peptides, the peptide via an ester bond with a Solid carrier is built up linked and after the assembly is separated from the solid carrier which according to the invention is characterized in that the separation of the peptide attached to the carrier is carried out in this way

909 83-4 / 1 52 0 '.; ■■

_ ₅ _

1904021

assumes that this is done with a mixture of an alcohol and an amine or quaternary ammonium compound of the general formula

respectively.

wherein R ₁ , R ₂ and R ^, either a hydrogen atom or an alkyl, cycloalkyl, aryl, aralkyl, alkaryl, amino-substituted or hydroxy-substituted alkyl group, at least one of the radicals R-, - R- , is different from hydrogen and R ₁ , R ₀ and R ₇ , can be the same or different, and where, when R ^, is hydrogen, R-, and R ₂ together are cyclic radicals of the formulas:

in which R denotes a hydrogen or halogen atom or an alkyl group, or R ,, common to the cyclic radical of the formula

and R, ge

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wherein R has the meaning given above, can represent, and IU, Rp., Rg and R "represent an alkyl, cycloalkyl, aryl, aralkyl, alkaryl, amino-substituted or hydroxy-substituted alkyl group, at least one of the radicals R ^ - R ₇ denotes an alkyl group and R ₂ , R ₁ -, Rg and R _{17 can be the} same or different, and where, when Rh and R ₇ each represent an alkyl group, .Rp. and Rg together cyclic residues of the formulas ''

in which R denotes a hydrogen or halogen atom or an alkyl group, or else, if Rh denotes an alkyl group, R _r , Rg and R ₇ together represent the cyclic radical of the formula

wherein R has the meaning given above, can represent, and X is a chlorine, bromine or iodine anion, brings into contact, dissolves the peptide therein and from the thereby

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resulting solution the peptide, optionally after previous purification, wins while the undissolved the remaining carrier is separated from the solution and, if desired, worked up.

In the method according to the invention, preferably in the Amines used as a mixture contain as alkyl radicals R ^, Rg and R, advantageously lower alkyl radicals, as such aryl radicals, preferably phenyl, and as sol .mino- or hydroxy- substituted alkyl radicals preferably also lower alkyl radicals.

If amine compounds are used in which R 1 and R p together form a cyclic radical, used amine compounds of the following structural formulas: (b)

R-

wherein jedfi'der R radicals either for a hydrogen or a halogen atom, preferably a chlorine atom, or an alkyl group,

wherein each of the R radicals has the meaning given above,

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-βία)

wherein each of the Η radicals also has the meaning given above has, and

where each of the E-ReBfee also has the meaning given above »

If one uses such an atom compound in which the radicals R ₁ * Rg and R, common «form a cyclic He®t, this formula has dt« n & ßhs dividing
(e)

vT

wherein each of the R-Reete also has the meaning given above
Examples of such amine compounds are triethanolamine,

Diethylamine * methylamine «dirnethylamine ^ -ethylene-

9 © 3 / i / Ii

hexylamine, pyridine, piperidine, pyrrolidine, pyrroline, pyrrole, α-, ß- and γ-picolines.

Provided that one quaternary ammonium compounds of the structural formula

(a) R ₅

¹ +

uses, in which the radicals R, - and Rg together have a form a cyclic radical, compounds of the following are used Structural formula

wherein ^ each of the R radicals is either a hydrogen or a halogen atom *, preferably a chlorine atom, or also a Alkyl group, Rg and% mean an alkyl group and X has the meaning given above, (o) "

A ^r

where R, Rg, Rg and X are defined as given above,

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1804011

ft

wherein R, Rg, Rg and X likewise have the meaning given above have and

where also each of the best B, Rg, has given meaning «
If- "one has such a qua ternary e
«• did« in which the Heat © lL «form Hg and R-ofoiiseohen remainder», has this
de formula,

and X oils above

wherein each of the radicals R, Rg and X has the meaning given above _e

·· 11 -

Monohydric or polyhydric saturated alcohols can be used as alcohol. Generally used? Solve either a hydrocarbon alcohol or an Ötltör- bssW * tiiiro-substituted hydrocarbon alcohol, old examples called selenium t aromatic hydrocarbon alcohols, for example benzyl alcohol, phenol, and the like

same! ftliphitische hydrocarbon alcohols, such as examples \ Bpiel8welse yinylalkohol, the lower alkanols (methanol, ethanol, propanol, isoamyl alcohol, etc.) "glycerol, ethylene glycol and the like, a cycloaliphatic alcohol such as cyclohexanol and the like, and their nitro and chlorine -substituted derivatives such as p-nitrophenol, chloral hydrate, and the like. The following alcohols are particularly preferred: benzyl alcohol, p-nitrophenol and the lower alkanols. If the alcohol used is a compound that is solid at normal temperature, such as p-nitrophenol, then this alcohol is expediently used in the form of a solution in a suitable solvent, such as a chlorinated hydrocarbon such as chloroform

The peptide adhering to the solid support is expediently brought to temperatures which do not exceed about 80 ° C., and generally at temperatures between about 20 and about 60 ° C., with the mixture of an alcohol and an amine or ouaternary ammonium compound in contact. You can also work at temperatures * higher than 80 C » υ.-άυά one separates the peptide from the carrier

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want, but it can happen that with such higher Temperatures a certain degradation of the 'peptide chain takes place, so that it is useful to avoid higher temperatures. The contact time must be sufficient to remove the peptide as completely as possible from the carrier material, and each required time varies depending on the working conditions used in the individual case. In general A contact time of between about 5 and 110 hours is sufficient to completely separate the peptide from the support material to reach.

In the mixture with which one adheres to the carrier material After bringing the peptide into contact, one uses such an amount of alcohol that is sufficient to dissolve the peptide it is separated from the carrier material; Excess amounts of alcohol have no adverse effect, one can Although smaller amounts of alcohol can also be used, there is then the risk that you will not get the maximum possible Releases amounts of peptide from the carrier material. The amine or quaternary ammonium compound used in the mixture is advantageously chosen in terms of quantity so that there is a ratio of about 5 to about 100 mM of amine or quaternary Ammonium compound per 1 mM "of peptide, preferably a ratio of about 10 to about 40 mM of amine or quaternary Ammonium compound per 1 mM of peptide results.

With the method according to the invention it is possible to a great extent advantageous and very effective way in a plague phase

Synthesis method to remove peptides obtained adhering to a solid support from the solid support. It will assumed that the mechanism of separation takes the form of a transesterification reaction, with the alcohol against the Solid carrier is replaced. The released In the presence of the amine, peptide is soluble in the alcohol and consequently can be easily removed from the solid support split off.

example 1

10 g chloromethylpolysbyrene-2 ^ divinylbenzene copolymer (1.84 mM Cl'Vs) were mixed with 150 ml of absolute alcohol, the Contained 5.9 S Boc-nitroarginine and 2.55 nil triethylamine, mixed. The mixture was refluxed for 48 hours at 80 ° C. with stirring by means of a magnetic stirrer. ■

Then the product was filtered and it was with absolute Washed alcohol, water and methanol. The product, Boc-nitroarginine linked to the polymer via an ester bond, was obtained in an amount of 11.8 g.

A 0.5 g sample of the esterified resin obtained as described above was in a mixture of 24.5 ml absolute Suspended methanol and 0.5 ml (j5.65 mM) triethylamine. This sample was stirred for 16 hours. The nitroarginine solution (separated from the resin) was filtered from the resin

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trated, and the pH of the solution was adjusted to 5 with acetic acid. The precipitated nitroarginine was: filtered off and washed with water. The amount of nitroarginine released, as determined by ultraviolet absorption, was 105 mg or Oj472 mnM / g of rosin ester. This represents an 80 # Yield based on a theoretical substitution of 0.59 mM / g rosin ester as determined by automated amino acid analysis. ./:-■

Example 2

A 0.1 g sample of that obtained as described in Example 1 Nitroarginine-esterified resin was made in a mixture Suspended 4.0 ml of absolute ethanol and 0.1 ml (0.75 mM) of triethylamine. This sample was stirred at 45 ° C for 88 hours. The nitroarginine was isolated as indicated above. An amount of 10 mg or 0.457 mM / g of resin was added be separated.

- ( ---.-V-.:-*: ■ - 'Example 3..

2 ml (10 raM) of .dicyclohexylamine were in 25 ml x anhydrous Dissolved methanol. To. 520 mg of resin esterified with nitroarginine (0.56 mM nitroarginine / g resin) were determined from this solution by amino acid analysis) added, after over After stirring overnight at room temperature, the resin became of the. Solution filtered off. After that, the resin was used twice Washed 10 ml of methanol. The amount of nitroarginine separated was 0.53 mM / g resin. This means a 59 ^ 1ge split after 16 hours.

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Example 4

500 mg (2.9 mM) of diphenylamine were in 25 ml of anhydrous Methanol 'dissolved »485 mg of nitroarginine were added to this solution esterified resin, as described in Example 3, added. The mixture was then left at room temperature for 24 hours touched. The resin was filtered off and the filtrate was evaporated to dryness. The residue was in 10 ml of 6 η hydrochloric acid dissolved and in an automated Amino acid analyzer analyzed. The amount of nitroarginine released was 0.4 mM / g resin, which corresponds to a 72 # spin-off.

Example 5

A 0.5 g sample of that obtained as described in Example J5 resin esterified with nitroarginine was dissolved in 1.5 ml (18.6 mM) of pyridine which had been diluted to 25 ml with methanol, suspended. The sample was stirred for 16 hours at room temperature. A 10 ml aliquot became dry evaporated and then in 10 ml of 6 η hydrochloric acid dissolved and analyzed in an automatic amino acid analyzer. It has been found that a lot of Nitroarginine of 0.027 mM / 0.5 g had been released.

The above example was made using piperidine, pyrrolidine, pyrroline, pyrrole and α-, β- and γ-picoline instead of pyridine with the same molar proportions as for pyridine. The results were practical in all cases the same.

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Example 6

A 0.5 g sample of the obtained as described in Example J5, Resin esterified with nitroarginine was dissolved in 25 ml of methanol containing 0.5 g of the quaternary hyamine 10-X (diehloride monohydrate) contained, suspended. The sample was stirred at room temperature for 16 hours · The amount of nitroarginine released was 0.020 as determined by ultraviolet absorption mM / g of rosin ester.

Example 7

56 g (400 mM) of p-nitrophenol were dissolved in 175 ml of chloroform which contained 50 ml (144 mM) of triethylamine, and this solution was added to 10 g of nitroarginine harvester as described in Example 3. The mixture was stirred at 6O ⁰ C for 20 hours, Ie was taken 5 ml of a sample and 10 ml of a β η hydrochloric acid extracted. The aqueous layer was separated off and analyzed analytically for amino acids. "The amount of nitroarginine released was 0.253 mM nitroarginine / 10 g resin ester"

The above example was repeated, and was thereby Benzyl alcohol tew, phenol or * -cyclohexanol used instead of p-Nltrophenol dissolved in chloroform obtained the same results «When using benzyl alcohol I found the further advantage that the. at the functional ester group remaining in the feptide chain easily remove bright »

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A particular ^Tr ortail of the inventive method is that in the previously described transesterification reaction, an active hydroxyl functional group remains on the resin, thereby being reused without further processing is possible "The following example illustrates this feature is the method according to the invention,

Example 8

A 5 g sample of nitrocellulose resin was suspended by suspending in 250 ml of a 10 # solution of triethylamine in methanol and stirring at room temperature overnight · It were 0.486 mM nitroarginine per g resin or a total of 2.43 mM Nitroarginine won. The resin was filtered and washed with dimethylformamide and methanol until it was free from Ultraviolet absorbing substances. After washing with methylene chloride the resin was treated with 1.6 g (7.29 mM or a 3-fold excess) of Boc-valine and 1.8 g of dicyclohexylcarbodiimide (8.7 mM or a 3.5-fold excess) in 60 ml of methylene chloride overnight with stirring at room temperature implemented * The analysis of the product for amino acids resulted in 0.231 mM valine / g resin

The boo-valine resin was further added with 1.2 g (7.29 mM) of Boc-olycin and 1.8 g of dicyolohexylcarbodiiraide (8.7 mM) in 60 ml of methylene chloride overnight with stirring at room temperature reacted · The rosin ester was washed with methylene chloride and acetic acid until it was free of acylurea · From the Amino acids on the resin became the Sohutz groups (Boc groups)

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removed with 50 ml of 1.5 HCl in acetic acid and washed with% acetic acid and then with methylene chloride to remove the chloride.

The rosin ester was reacted in the same way with 1 _S 4 g (7 # 29 vM) Boc-alanine and 1.8 g (8 * 7 mM) dicyclohexylcarbodiimide in 60 ml methylene chloride overnight with stirring at room temperature »A hydrolyzed sample of the product , which was subjected to an analysis for amino acid, gave the following valuesι 0.171 micromoles of valine, 0.216 micromoles of glycine oil, 0.39 ¹ J micromoles of alanine.

It can be seen from Example 8 that the following processes took place havei

a) Nitroargine became almost quantitative by transesterification away from the Harz *

b) During the transesterification, active hydroxyl groups remain the resin.

0) In the conduction mediated by DloyolohexylcarbodiiraW « aation of Boc-Valine with the resin succeeded in making the Boc-VaXin »

eeter of the resin to form.

d) In the case of a second condensation resulting in CtXyein has succeeded in forming a beneficial glycine »

Valln-Harseetera the remaining ^ '3 $ uvs: tjlgvvi $ p <dn - to link to resin,
e) In the connection, the condensation with lignin is carried out "

After removal of the protective groups, the dipeptides valjäralanin were found and Glycylalanih are formed,

From the above description it can be seen that the inventive Method in general for separating and releasing a peptide which is attached to a solid support material Is linked via an ester bond at the terminal C, is suitable, and therefore that is according to the invention The method in no way refers to the carrier materials and peptides specifically mentioned for example purposes only limited «For example, you can use any Use any solid substance, provided that the two previously mentioned, for the solid phase synthesis for production properties required by peptides, i.e. inert behavior towards the reagents used and large active surface area as well as one to form an ester bond with the terminal carbon atom of the peptide has a suitable group, are examples of such carrier substances a hydroxymethyl substituted styrene-vinylbenzene copolymer or any polymer that has free hydroxyl or chloride groups »Further, any of a great variety of amino acids as the portion of the Peptide chain via which the C-terminal ester bond with the carrier material takes place, use. Examples of such Amino acids are alanine, arginine, aspartic acid, asparagine, Cysteine, cystine, diiodotyrosine, glutamic acid, glutamine, glycine, histidine, hydroxyproline, isoleucine, leucine, lysine, methionine, Norleucine, ornithine, phenylalanine, proline, serine, threonine,

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Tryptophan, tyrosine, valine, and their substituted derivatives in common use, such as Fitroarginin · So were "B # z, the following peptides from a solid support removed to further illustrate the present invention, ·

Example 9

A 0.5 g sample of a deeapeptide resin ester (L-Mitroarginyl-L-leucyl-Ir-valyl-Ir-iraldazolbenzyl-L-histidyl-L-isoleucylepsilon-carbobenzoxy-L-lysyl-Ir-inethionyl-Ir'tryptophyl-L -threonyl-Ir-phenyiaianin), which had been prepared according to the general procedure of Merrifield (described 1963 - J, Asu Cheia. βοο ·, 85, 21 ^ 9), was suspended in 23 ml of absolute methanol and 2 ml of diethylamine and Stirred at 45 ° C for 60 hours · The amount of released peptide, determined by UXtraviolet absorption of the nitroarginine, was 0.195 mM / g · In the amino acid analysis of an acid hydrolysis acid of the decapeptide given above, 0.268 raM / g of nitroarginine was released ·

A 0.21 g sample of dipeptide (mitroarginyl-leucine) - resin ester was suspended in 9 * 0 ml of absolute methanol, ϋ., 8 ml of Cfiloroforra and 0.2 ml of triethylamine and 16 StuiideKiL long, at 25 ^: ß The amount of released Mitroarginine amount, determined ultravloXett absorption, 0.3 ^ 6 wM / g * Another sample of this material is feydrolysieai; % with an itaxBosäwre analysis. " It wurüen 0.3 ^ 2 υϋξ / g Mifcroarginln andl 0, ^ 2 tsm / g, Xeucin determined ·

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Example 11

A 0.5 g sample of deeapeptide resin ester was dissolved in 23 ml of absolute Ethanol and 2 ml of triethylamine suspended and stirred at 80 ° C for 110 hours. The amount of released The peptide was 0.157 m / g as determined by the alu violet absorption of nitroarginine.

Another important technical step that is achieved with the method according to the invention is that a functional ester group remains on the peptide during the transesterification reaction described above, which makes it possible to separate the peptides with a lower molecular weight from the peptides with a higher molecular weight. For example, in the production of a decapeptide, peptides with a lower molecular weight are also formed, and during the separation of the peptide, such peptides with a lower molecular weight also dissolve in the presence of the amine, together with the decapeptide, in the alcohol. The peptides with the lower molecular weight, if they contain ester-functional groups, are soluble in conventional solvents, for example alcohols, e.g. the aforementioned alcohols, acetone, tetrahydrofuran and the like, while the peptides with higher molecular weight, which contain ester-functional groups, are soluble in such solvents not soluble sin * i. Accordingly, the peptide solution obtained in the process according to the invention can be evaporated to dryness and the solid residue can be treated with a suitable solvent, in which case the peptides with the lower molecular weight

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be dissolved in the solvent. In this way, the peptides with a higher molecular weight are obtained in proportions with a lower one Free form molecular weights. This is illustrated in the following example

Example 12

In a suitable container equipped with a condenser, thermometer, heater and stirrer, 120 g of a decapeptide resin ester (L-nitroarginyl-Lleucyl-Lvalyl-imidazolbenzyl-L-histidyl-L-isoleucylepsilon-carbobenzoxy-L -lysyl-L-methionyl-L-trypophyl-L-phenylalanyl-L-threonine), prepared according to the general procedure of Merrifield, described 196 ^ - J Am. Chem. Soc "85". 5520 ml of absolute methanol and 480 ml of triethylamine were added to this. This suspension was stirred at 50 ° C. until no further increase in the Utraviolffitt absorption, determined for nitroarginine (extinction coefficient 17.3 · ^ at 268 myu) was found. The piltrate was filtered hot from the resin and concentrated to dryness under reduced pressure. Three ultraviolet spots and five ninhydrin positive spots were found on paper electrophoresis (Whatman 5MM paper) at pH 1.7. The residue was triturated with methanol several times until a uniform electrophoretic pattern was obtained, which is an indication of the uniform composition of the residue.

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In the above example, the one ester function was used Group having peptide as a product of the here • ahriebentn However, peptide separation can also be obtained Separate piptides released by other methods in the same way.For example, a peptide, which by means of using hydrogen bromide and Trifluoroacetic acid working method has been released, a with an alcohol in order to create an ester-functional Group, and then treat in the manner previously described to the fractions with lower molecular weight to separate.

The method according to the invention has a number of important advantages. Relatively mild reaction conditions have to be used and a degradation of the peptide chain can be prevented. Furthermore, in the method according to the invention, the amount of peptide separated from the carrier material is considerably increased compared to previously known methods of this type. In addition, in the process according to the invention, there is no need for chemicals that are harmful to health and difficult to handle, such as hydrobronic acid and trifluoroacetic acid; and in general the method according to the invention is very much simpler, so that it can be more easily controlled automatically. In addition, falls when ssan p-Witrophenol used as the alcohol in the Abtrenrastuffe, the peptide in the form of an activated ester capable of linking to üeu ^ eam directly with an on another carrier material smhaf temfei peptide chain or an amino acid.

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Another advantage of the method according to the invention is that the carrier material, of from which the peptide has been separated, an active hydroxyl functional Having group that enables the reuse of the carrier material without further treatment. Advantageous is also that one has an ester functional group containing recovered peptides in a simple manner in fractions with higher and those with lower molecular weights can sever.

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Claims (10)

- 25 claims 1. Solid phase synthesis process for the production of peptides, wherein the peptide has an ester bond with a Solid carrier linked built up and after the build up is separated chemically from the solid carrier, characterized in that the separation of the adhering to the carrier Peptide makes in such a way that this with a mixture of an alcohol and an amine or quaternary Ammonium compound of the general formula fc ₊ - R ₂ -N- R ₁ ^or R4 - N - R ₆ χ ι t ■ wherein R ₁ , R ₂ and IW are either a hydrogen atom or an alkyl, cycloalkyl, aryl, aralkyl, alkaryl, amino-substituted or hydroxy-substituted alkyl group, at least one of the radicals R ₁ - R, different from hydrogen and R ₁ , Rg and R, can be the same or different, and where, when R, is hydrogen, R ₁ and Rg together are cyclic radicals of the formulas: :O: R-f T-R pP ^ v x ^ -R in which R denotes a hydrogen or halogen atom or an alkyl group, or also R ₁ , R ₂ and R, 909-834 / 1520 common to the cyclic radical of the formula ft "" N ^ £ j (r * ~ -m »x \ wherein R has the meaning given above, can represent, and R ^, R, -, Rg and R "for an alkyl, cycloalkyl, Aryl, aralkyl, alkaryl, amino substituted or hydroxy substituted Alkyl group, at least one of the radicals R ^ - R "denotes an alkyl group and Rj ,, Rj-, Rg and R" may be the same or different, where, if Rj. and Ry each stand for an alkyl group, R, - and Rg together cyclic residues of the formulas "XX in which R denotes a hydrogen or halogen atom or an alkyl group, or, if R ^ denotes an alkyl group, R ₅₅ Rg and R "together represent the cyclic radical of the formula R .1 wherein R has the meaning given above, can represent, and X is a chlorine, bromine or iodine anion, in Brings contact, dissolves the peptide in and out of it 909834/1520 resulting solution the peptide, optionally after previous purification, wins while the undissolved the remaining carrier is separated from the solution and, if desired, worked up. 2. The method according to claim 1, characterized in that it does not carry out the in-contacting at about 8O ⁰ C for rising temperature. 3. The method according to claim 1 or 2, characterized in that that the amine or quaternary ammonium compound in a ratio between about 5 to 100 Millimole of amine or ammonium compound is used per millimole of peptide resulting amount. 4. The method according to claim 1 to 3 *, characterized in that that one for the separation consists of a chloromethylated styrene-divinylbenzene copolymer Carrier attached peptide used. 5. The method according to claim 1 to 4, characterized in that that the alcohol in the mixture is hydrocarbon alcohols which are nitro- or chlorine-substituted can, preferably lower alcohols, used. 6. The method according to claim 4 or 5 » characterized in that such a mixture is used which contains p-nitrophenol or benzyl alcohol as the alcohol and an alkylamine, preferably triethylamine, as the amine compound. 7. The method according to claim 1 to 6, characterized in that the peptide dissolved in the mixture by evaporating the solution to dryness in solid form brings and then cleans it in such a way that one with a solvent, preferably an alcohol, the Dissolves fractions with a relatively lower molecular weight. 909834/1520 8. The method according to claim 4 to 7 > characterized in that a decapeptide is obtained when the solution is evaporated to dryness, and the solvent dissolves peptide tractions with this relative to lower molecular weight. 9. The method according to claim 4 to ¹ J, characterized. draws that a mixture of methanol and triethylamine is used, the decapeptide L-nitroarginyl-L-leucyl-L-valyl-imidazole-benzyl-L-histidyl-L-isoleucyl-epsilon-carbobenzoxy-L-lysyl from the resulting solution -L-methionyl-L-tryptophyl-L-phenylalanyl-L-threonine is obtained in solid form and the lower molecular weight components are extracted from it with methanol as the solvent. 10. The method according to claim 1 to 9 *, characterized in that the working up of the solution separated carrier in the "way that you bring this into contact with an amino acid and that !! Carrier material with the amino acid to one for further peptide synthesis esterified carrier substance having suitable ester bond. 909834/1520