DE2032329A1 - See new partially protected basic amino acids and peptides of ulcer-inhibiting effect and processes for their production - Google Patents

Description

Dr. rer.nat. Bruno Reitzner

Patent attorneys ·

8 Munich 2, Hermann-Sack-Str. 2

June 29, 1970 Our sign P-3788-1

Patent to m'e 1 thing for

Judge Gedeon Vegyeszeti Gyar Rt., Budapest X, Hungary

concerning

New partially protected basic amino acids and peptides of ulcer inhibitory effects and process for their preparation

The invention "relates to the manufacture of new, partial protected, basic level (apart from the -amino group containing another basic group) amino acids and peptides, the show an ulcer-inhibiting effect

Recently, more and more references are appearing on the biosynthesis of acidic mucopolysaccharide protein -Complexes, as well as about the different ones, in the connective tissue- -Diseases play a significant role enzyme systems (ζ · Β · elastase, pronase, collagenase etc ·) · den in the connective tissue cells The various proteolytic enzymes that are present have recently also been assigned an important role in inflammatory diseases. The inhibition or stimulation of these Enzymes Form a New Direction in Drug Research · There have been various drug preparations recently introduced into practice which as inhibitors of certain Enzyme systems act (ζ · Β · MonoaminooxydaGe inhibitors, trypsin " or kallikrein inhibitors, etc.)

^67/81 * "· 00Ü88S / 2249

It was now in surprising wqice pounds, in addition to the partially (each protects on baciochon alaino acids of the general formula I.

Ac - X - OH - COOR

L ₂ . ν

wherein X is an organic containing an acylatable nitrogen atom Group, Ac bonded for oine sum mentioned Sticketoffatom Acyl group and R for hydrogen or for an alkyl group of 1 up to 5 carbon atoms, as well as their pharmaceutically applicable ones Acids formed addition salts beneficial pharmacologicche Properties, especially an ulcushcmi'.ende effect, also promoting the incorporation of sulfur into the cartilage tissues and show body weight reducing effects

Bio new compounds inhibit practically completely in rats the development of artificially induced ulcers. It can it is assumed that this effect is involved with the stimulation dec or in the biosynthesis of the acidic mucopolyaccharides Enzyme or enzymes in connection with ctoht. With this Effect is probably also related to the experimentally proven fact that the new compounds incorporate from S * - ^ into the cartilage and stomach of the rats increase; the incorporation can be considered a Indicator of the biosynthesis of the acidic! Lucopolysacch & rid protein -Complex can be considered · It can be found in the literature so far no clues are found «after soft a substantial Increasing the fixation of s "in the cartilage in vivo experimentally would have been established · The mechanism of the above-mentioned appetite-reducing effect cannot be compared to that of the already common appetite suppressants, since an effect on the neutral nervous system has not been proven with the new compounds could be

Purely the novel compounds according to the invention of the general Formula I is the presence of the free ctf amino group characteristic, which ensures the water solubility of these new compounds.

In the place of X, practically any, an acylated ^ organic groups containing a free nitrogen atom, e.g. an in alkylene chain substituted by an amino radical in the O (* position (as with lysine), one substituted by a guanidino oct

009885/2249

BATH

(as with arginine) or an alkylene group substituted by a heterocyclic radical containing the acylatable i> tickctoriatom & le Rinc member (ζ The acyl group of an aliphatic carboxylic acid of 1 to 5 carbon atoms or a carboxylic acid containing a phenyl group, such as a benzyloxycarbonyl group, an acyl group containing more basic groups, i.e. an amino acyl group} in the latter case, the amino group of this acyl group can be advantageous also in a manner known per se, s, B · be protected by a p- -OhlorbeneyloxycarbonylreEt ^ eBChichert · The substituent R can be hydrogen or an alkyl group of 1 to 5 carbon atoms - in the first case let the iaolekül to form an inner salt while lower alkyl ethers in the Lotzeren Pail | stand·

The new compounds of general formula I are exv manufactured according to the invention in such a way that one, another basic rect-containing A-amino acid of the general formula II

H-X-r CH-COOH

where X has the above meaning, in a manner known per se in a the et-amino group and the Carbojtylgruppe in protected Pro * containing metal complexes, advantageously copper complex derivative transferred, then dieces with a / a, sum introducing the in the final product desired acyl radical Ac suitable acylating agents, expedient with an acid chloride of the general formula Ac-Cl or with a derivative containing the carboxyl functions in activated formula acylated the acyl group Ac, then the Uetallkomplea: in known Way decomposed and desired trap the compound obtained general formula I, which instead of E is a hydrogen atom contains, alt a sub-introduction of an alkyl group from 1 to 5 Carbon atoms suitable means esterified and / or the product Converted into a therapeutically applicable salt

Is lore in as a starting material of the general formula II used, then the erfindungßgemäse.e method can especially are advantageously carried out in such a way that a copper (II) -

BATH ORIGINAL

U O £ O

-Sals used to form the metal complex and the S-amino group of the lysiiss was acylated with p-chlorocarbons in the presence of acid binders, expediently by magnesium oxide. In this way, after decomposition of the metal complex, a compound of the general formula I is obtained in excellent yield, in which the acyl group Ac is replaced by a p-chlorobenzyloxycarbonyl group _} X by a © -KH-OH ₂ ^ Hp-CHp-CH ₂ group and E is represented by a hydrogen atom. This compound can then be converted into the corresponding ester in a manner known per se, for example by simultaneous reaction with thionyl chloride and methyl alcohol »

The compounds of the general types which can be prepared according to the invention Formula I are new products not previously described in the literature

The compounds prepared according to the invention of the general Formula I are applied depending on the Reaction conditions in the form of free bases or acid addition salts obtained · Compounds of the formula I, which at the place of R contain a hydrogen atom, can also Metal salts, or «» ammonium or amine salts, form · From the in compounds obtained in the form of salts, the bases or basic acids can be released in a manner known per se, or Such compounds can be converted into other desired, expediently therapeutically applicable salts, especially acid addition salts transferred v / earth

The method according to the invention is carried out by the following Examples illustrated in more detail.

Example 1:

2- (p-chlorobenzyloxycarbonyl) -L-lysine-jne thyle st er .HCl ·

a) 9.0 g (0.04-9 M) L-lysine monohydrochloride are dissolved in 100 ml of water, 3.5 g of copper carbonate are added to the solution and the suspension obtained is boiled for half an hour. After cooling to room temperature the reaction mixture is filtered off. 6.0 g of magnesium oxide are added to the titan blue filtrate, the mixture is ^{cooled to 0} ° C. and, at this temperature, ₉ with stirring, within half an hour with 12.7 ml of p-Cfalorbenzyloaycarbonylchlorid (cf. L. Klsfaludy and S. Dualszky » Acta Chira · Hung «2J. 501, 1960) added drop by drop. The reaction table

009885/2249

is stirred for a further 2 hours at room temperature, then ^{cooled again to 0 0} O and the precipitated lavender blue precipitate is filtered off, washed thoroughly on the filter with water, then with alcohol and finally with ether and dried in the air , 6 g copper complex obtained

The copper complex obtained in this way is thoroughly powdered and suspended in 325 ml IT hydrochloric acid. Hydrogen sulfide gas is passed through the suspension for 2-3 hours, then the suspension is boiled and filtered in a hot state to remove the copper sulfide. The water-clear filtrate is cooled and treated with conc. Ammonium hydroxide solution adjusted to pH = 5, the weta precipitate obtained is filtered off and washed with water Wash and dry in a vacuum desiccator over phosphorus pentoxide 11.9 g of t-Cp-chlorobenzyloxycarbonyl-L-lysine (77 / Sd. Th) are obtained * f. 236-238 ⁰ O.

b) 6.3 g (0.02 M) of E- (p-chlorobenzyloxycarbonyl) -Ir-lysine are suspended in 42 ml of absolute methanol in a four-necked round bottom flask equipped with a stirrer, thermometer, dropping funnel and G CaOlg tube suspension is cooled to ⁰ ° C and added under stirring with 1.7 ml (0.023 M) of thionyl chloride dropwise · After a few minutes will obtain a clear yellow solution, this will be one and a half hours at 40 ⁰ O, then 20 hours at room temperature calmly. The solution is then filtered off and evaporated in vacuo. The yellowish-white moist residue is dissolved in methanol and the solution is evaporated to dryness again. This i | Operation will be Z ¹ O) imal repeated and finally the product from methanol / ether is crystallized. The mixture is left to stand for a few hours in the refrigerator, then the crystals are filtered off, washed with a mixture of Ither and methanol (3 * 1) and dried in a vacuum desiccator over sodium hydroxide. There are 5 lg _f £ - (p-Ohlorbenzyloaycarbonyl) -L-lysine methyl ester hydrochloride, mp .150-160 ⁰ O. R _f = 0.48 (the dünnschicbtchromatographische study was performed on silica gel • for steel · Plate ethyl acetate-pyridine-glacial acetic acid-water 60x20t6ill carried out! The development took place with ninhydrin or with chlorine and toluidine.

009 885/2249

Analyst! calculated! C 49.3 #, HS ₈ I found! O $ 4, 5 % _t H.

Example 2ι

a) 16.9 S (O ₁ IM) L - <) vnifchi & ° @ enohydxoehlorid be la

170 ml of water, the solution ai _e ©% ^ 5 g lnpfesearboaat v © r st set and the Susptnsloa obtained is lös to Si @ d®a heated compartment cooling, the Gasiseh is filtered _D dark blue. Filtrate is cooled to ⁰ ° C uad oatsr Rütadfi tso.pfen \ 7oise ₀ 'equilateral within 20 Minatöai ai * SO ^ ₀ oxycarbonyl chloride and 101.4 ml of 21? Sodium hydroxide solution added ö The R-3aktionegemiech is still ela © femllb @ S ^ öaS © "Öoi gorUhrt, then the easgeiB © iii © iiiB, © light blue precipitate a ^{fe <co} filtered, with water and ethanol g © waseli®a uad ' m air dried

The obtained Kupferkompl®2 £ visä_ Ib ^ QO saspendiert ¹ ^ l water and ScnwefelwasserstoffgaB is lüsg 2-3 Stmiea · guided into the suspension, the mixture is in good ^ eseefeloseoa® "Gofass üb ©? Hacht left to stand "Aa next day, the suspicion is diluted with 1000 ml of water and until the boil a © Alt gto The hot mixture is mixed with as much saleattje © v" s © tafe _B that the nebea the copper (H ) -ßulfid ausgeschieflene wise Hied © »schla @ again Boll · dissolve wad filtered the Kupfensulfid wird.toiss the Piltrat is provided after the AbkÜhlea äerok oils Zogab © το & cons · Hamoniumhydroxydlösung to pH a 4-5 · bite precipitated white product is Filtered off, washed with water and i® Ya & uumexsiccator, dried over phosphorus pentoxide · Ss warden 17 © β S 5 »(p» chlorobenzyloxyoarbonyl) -Ir.ornithine arhalteag Γ · (W) -255-255 ° C.

b) In a _{Tierhale-Himdkolbeii equipped with Eührer t} TaermoiaoteE © drip funnel uad Ohlorkalaiuarohrub 15 »! (0.05 M) • - (p-OhlorbeneyloxycarbonfD-L-oEnithiÄ suspended in 105 ml absolute ethanol. Pie suspension is ^{cooled to 0} ° C. and, with stirring, old 4.25 μl (0.0575 μl) thionyl chloride drop by drop "Bach versetst agree · * minutes will have a clear, yellowish solution for 1.5 hours at 40 ⁰ O and then 20 hours at Siaaertemperatiur is allowed to stand is · The solution then f il-

009835/2249

tricrfc and vardarcpft to dryness in Valiuun · The yellowish white solid residue is dissolved in methanol and the solution evaporated again · This operation is repeated twice and then the residue is crystallized from a mixture of methanol and Ither. The crude product (13.4 g) is again uokrictalliciert from Mothanol / Ithcr. 9.3 g of Ä- (p- -Chlorbenzyloxycnrbonyl) -L-ornithine-ethyl-8ter hydrochloride are obtained at P, 160-162 ⁰ Cj R _f = 0.43,

Analysisι C ₁₄ H ₂₂ O ^ N ₂ Cl ₂ (351.22)

calculated: C 4-7.9 % $ H 5% 1 ft, N 8.0 JS, Cl-20.2 % \ found! C 47.9 H 5.9 %%% t 8.1% _K% Cl 20.3% * Example 3i

^ - (p-CbJ.orbenayloxycarbonyl-L-tyrosyD-iclysine · "

In a üit stirrer, Thcrnomcter, dropping funnel and calcium chloride tube cuccestattcten 250 nl VierhalE-Eunalcolben v / ground 10t3 C (29 | 4 mil) p-ChlorbDnsyloxycarbonj'l-L-tyrosine ml abs · tetrahydrofuran elöct ^ _t depart in 100; previously with ^, 1 ml (29.4 Triethylamine was vorcetzt. The solution is cooled to -15 ⁰ C and un ^ or stirring with 3.82 al ■ (29.4 "Μ) ChlorkohlenBäure-isobutylecter dropwise added with such GoschwindiGkeit that the temperature dos mixture be kept below -10 ⁰ C to · Haoh the completion of the dropping, stirring is continued, the obtained welsee Suepension at -10 ⁰ C for a further 10 minutes · Then, an aqueous solution of the Kupferkonplexes is added dropwise which Were you prepared in the following manner the suspension of 5 _f 5 6 (30 mM) Ii -Lysine fflonohydrochloride and 1.75 S copper corbonate in 30 ml of water are boiled for half an hour | After cooling, the mixture is filtered and the dark blue filtrate is mixed with 15 ml of 2N sodium hydroxide solution.

During the dropping of the above wiser solution is the temperature of the reaction mixtures β at -10 ⁰ C held · After the dropwise addition, the cooling bath is removed and the Reaktlonegemisch to warm up to room temperature stirred The light blue suspension is in 600 nl Lasser poured · After 1 Hour cooling, the lavender blue Hiederecblac is filtered off, washed with water and air-dried. Yield 11-12 ge

The Dipe ^ tidkupferkouple obtained in this way ?: v.ird

suspended in 50 al of water and the suspension with 30 ml of N

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BATH ORIGINAL

Hydrochloric acid added »Build, is unties? Heating of hydrogen sulphide gas Passed into the suspension for 5 hours and the resulting lupfer (II) sulfide is filtered off through the holes © Hack dem The filtrate is cooled by adding sodium hydroxide solution pH β 5 set · The precipitated white product is filtered off and old water washed

3s 6.15 g of S-Cjp-Chlorbsnayl -L-lysine are obtainedj P © 218-220 ° (Zers®) § H _f β 0 _s 2

Analysis »C ₂₅ H ₂₈ O ₆ I ₅ Ol (477 · 93) calculated 1 C 57.8 JS ₉ Ξ ° 5 _Θ 9 ff» "H β ₉ Β % ₉ 01 7» 4 ^ g found!. 01 7, 4 % *

88B5./2249

:. "· ORIGINAL INSPECTED

Claims (1)

Patent claims: (L>) Part ie II protected basic amino acids or -amino-. ' acid esters of the general formula I '■ Ac - X - OH - OOÖR wherein X is one containing an acylatable nitrogen atom organic group, Ac for one bonded to the nitrogen atom mentioned Acyl radical and R represents hydrogen or an alkyl group of 1 to 5 carbon atoms ", and their therapeutically applicable Salts. - 2 Process for the production of partially protected ' basic amino acids or amino acid esters of the general formula ι ■ ■ "\ · Ac-X-OH-COOR I ■. ' NH ₂ where X stands for an organic group containing an acylatable nitrogen atom, Ac for an acyl radical bonded to the nitrogen atom mentioned and R stands for hydrogen or an alkyl group of 1 to 5 carbon atoms, and of their acid addition salts or of the optionally internal salts of the free carboxylic acids, thereby characterized in that one, a further λ basic radical containing QC -amino acid of the general formula II H - τ - OH - COOH ■ -. _% ■ ' I (II) wherein X has the above meanings, in known manner in a o (-Aminogruppö and Oarboatylgruppe in protected form containing metal complex, copper complex derivative advantageously converted, "T a nn dieees with one, for the insertion of the desired Bndprodukt acyl Ac suitable acylating agent, expediently with an »acid chloride of the general Jorael Ac-Ol or with a die 009885/2249 Carboxyl function in activated form containing derivative of the acyl group acylated, then the meball complex la is decomposed in a known manner and, if desired, the compound of the general formula I obtained, which contains a hydrogen atom at Stell® of H, with an acyl group of 1 to 5 to introduce Carbon atoms suitable agent esterified and / or the product obtained, if desired, converted into a SaIs ₉ advantageously into a therapeutically applicable acid addition agent » 5. Traversing «! naea claim Z ₉ for the production of £ - (p- - ^ hlorbentyloxycarbonyl ^ lysiüi and leg esters "characterized in that lysine is used as the starting material of the formula II" which et S by reacting sea with a lupf er (U) -SaIz , in the corresponding lletallkoraplex derivative transferred _B, the C-amino -then with p-Chlorbenzylozycarbonylehlorid aeyliert _f the Metäll "komplez zweckmSeeig äiereetst with Schwefalwaeserstoff and the received product desired skins ₀ practical old; Help ELNES mixture of thionyl chloride and a IUohol of 1 up to 3 carbon atoms, converted into a lower alkyl ester 00983S / 2249