U.S. patent application number 14/848500 was filed with the patent office on 2016-02-04 for 2-amino-3-methyl-hex-5-enoic acid and its use in the prodction of peptides such as bacitracins.
The applicant listed for this patent is XELLIA PHARMACEUTICALS APS. Invention is credited to Vidar Bjornstad, Jon Efskind, Martin Mansson, Christine Senstad.
Application Number | 20160031798 14/848500 |
Document ID | / |
Family ID | 43066593 |
Filed Date | 2016-02-04 |
United States Patent
Application |
20160031798 |
Kind Code |
A1 |
Mansson; Martin ; et
al. |
February 4, 2016 |
2-AMINO-3-METHYL-HEX-5-ENOIC ACID AND ITS USE IN THE PRODCTION OF
PEPTIDES SUCH AS BACITRACINS
Abstract
The present invention concerns 2-amino-3-mmethyl-hex-5-enoic
acid, its use for the production of peptides such as bacitracins
and a method for producing it.
Inventors: |
Mansson; Martin; (Oslo,
NO) ; Senstad; Christine; (Hosle, NO) ;
Efskind; Jon; (Oslo, NO) ; Bjornstad; Vidar;
(Sorumsand, NO) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
XELLIA PHARMACEUTICALS APS |
COPENHAGEN |
|
DK |
|
|
Family ID: |
43066593 |
Appl. No.: |
14/848500 |
Filed: |
September 9, 2015 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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13502144 |
Apr 16, 2012 |
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PCT/EP10/64511 |
Sep 30, 2010 |
|
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14848500 |
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61255521 |
Oct 28, 2009 |
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Current U.S.
Class: |
514/561 ;
562/574 |
Current CPC
Class: |
C07K 7/58 20130101; C07C
229/08 20130101; C07C 229/30 20130101 |
International
Class: |
C07C 229/30 20060101
C07C229/30 |
Claims
1. A peptide comprising a 2-amino-3-methyl-hex-5-enoic acid residue
having the following side chain ##STR00009## wherein the peptide is
not a bacitracin.
2. A composition comprising 2-amino-3-methyl-hex-5-enoic acid.
3. The composition of claim 2, wherein the
2-amino-3-methyl-hex-5-enoic acid is in the (2S,3S)
configuration.
4. The composition of claim 2, wherein the
2-amino-3-methyl-hex-5-enoic acid is in the (2R,3S)
configuration.
5. The composition of claim 2, wherein the
2-amino-3-methyl-hex-5-enoic acid is in the (2S,3R)
configuration.
6. The composition of claim 2, wherein the
2-amino-3-methyl-hex-5-enoic acid is in the (2R,3R)
configuration
7. The composition of claim 2, in the form of a pharmaceutical
composition comprising a pharmaceutically acceptable excipient.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. Ser. No.
13/502,144 filed on Apr. 16, 2012 which is a 371 of PCT/EP10/64511
filed on Sep. 30, 2010 which claims the benefit of priority to U.S.
61/255,521 filed on Oct. 28, 2009, are incorporated by reference
herein in their entirety.
FIELD OF INVENTION
[0002] The present invention relates to new amino acid
compounds.
BACKGROUND OF INVENTION
[0003] The majority of proteins are made of 20 different
.alpha.-amino acids: Alanine, Arginine, Asparagine, Aspartic acid,
Cysteine, Glutamine, Glutamate, Phenytalanine, Glycine Histidine,
Isoleucine, Lysine, Leucine, Methionine, Proline, Serine,
Threonine, Tryptophan, Tyrosine and Valine.
[0004] However, there are many biologically active peptides
comprising other amino acids such as: Homocysteine, Ornithine,
4-Hydroxyproline, 5-Hydroxylysine, Selenocysteine, Formylmethionin,
Phosphoserine, Acetylserine, Methylarginine etc.
[0005] Amino acids in D-configuration are common in non-ribosomally
synthesized bacterial peptides and less common in ribosomally
synthesized proteins. For example in the non-ribosomally
synthesized Bacitracins, the amino acid residues in position 4, 7,
9 and 11 are usually in D-configuration (Glu, Orn, Phe and
Asp).
[0006] Bacitracins are peptide antibiotics naturally produced by
Bacillus subtilis and Bacillus licheniformis. Several Bacitracins
have been identified of which Bacitracin A is of primary importance
and is highly active (Biochemistry, vol. 39 no 14, 2000, page
4037-45 by Epperson and Ming). The primary structure of Bacitracin
A is
NH.sub.2-L-Ile.sub.1-L-Thiazoline.sub.2-L-Leu.sub.3-D-Glu.sub.4-L-Ile.sub-
.5-L-Lys.sub.6-D-Orn.sub.7-L-Ile.sub.8-D-Phe.sub.9-L-His.sub.10-
D-Asp.sub.11-L-Asn.sub.12-COOH which is cyclized between the
.epsilon.-amino group of L-Lys.sub.6 and the R-carboxyl group of
L-Asn.sub.12.
PRIOR ART
[0007] Several non-ribosomally synthesized peptides comprise
unusual amino acids. For example cyclosporin A comprises
2(S)-amino-3(10-hydroxy-4(R)-methyl-6(E)-octenoic acid which is
crucial for binding to the intracellular receptor for cyclophilin,
and thus for its immunosuppressive activity (Journal of Biological
Chemistry, vol. 268 no 35, 1993 by Offenzeller et al.).
[0008] Several uncommon amino acids resemble the structure of
Isoleucine:
[0009] 2-Amino-5-methyl-5-hexenoic acid, a new methionine analog,
was isolated from a fermentation broth of Streptomyces (Journal of
Antibiotics vol. 32 no. 11, page 1118-1124, 1979 by Takeuchi et
al).
[0010] 4 methylene-norleucine and 2-aminohept-6-enoic acid are
compounds with the formula: C.sub.7H.sub.13NO.sub.2.
[0011] 4 methyl-norleucine is an isoleucine derivative which can be
incorporated into a recombinant protein. (J Pharm Biomed Anal, vol
31. no. 5, 2003, page 979-987 by Muramatsu et al).
[0012] 2-amino-3-methyl-4-pentenoic acid is an unsaturated
isoleucine analogue which can be incorporated into proteins
(Chembiochem vol. 7 no. 1, 2006, page 83-87 by Mock et al).
[0013] The unsaturated norleucines of Amanita solitaria. Chemical
and pharmacological studies (Lloydia vol. 36 no. 2, 1973, page
69-73 by Chilton et al).
[0014] Beta-methylnorleucine, an antimetabolite produced by
Serratia marcescens (J Antibiot, vol. 34 no. 10, 1981 page 1278-82
by Sugiura et al)
[0015] U.S. Pat. No. 6,168,912 describes various all - derivatives
of amino acids for use in a multidimensional combinatorial chemical
library.
[0016] Allyiglycine and crotylglycine are described in o mat of
Bacteriology, vol. 148 no, 1, 1981 by Kunz et al.
SUMMARY OF THE INVENTION
[0017] The invention concerns the compound
2-amino-3-methyl-hex-5-enoic acid.
[0018] Although we use the name 2-amino-3-methyl-hex-5-enoic acid,
it covers the same compound as 2-amino-3-methyl-5-hexenoic
acid.
[0019] The structure of this amino acid compound in its free form
is represented by Formula I:
##STR00001##
[0020] Formula I is not to be interpreted as a Fischer projection,
but rather as a compound without specification of
stereochemistry.
[0021] The amino acid of Formula I comprises two chiral carbon
atoms. The present invention covers the four stereoisomers of
2-amino-3-methyl-hex-5-enoic acid:
[0022] (2S,3S)-2-amino-3-methyl-hex-5-enoicacid,
[0023] (2R,3S)-2-amino-3-methyl-hex-5-enoic acid,
[0024] (2S,3R)-2-amino-3-methyl-hex-5-enoic acid, and
[0025] (2R,3R)-2-amino-3-methyl-hex-5-enoic acid.
[0026] Further, the present invention covers salts and ions of
2-amino-3-methyl-hex-5-enoic acid.
[0027] Further, the present invention covers amino protected and
carboxy protected derivatives of 2-amino-3-methyl-hex-5-enoic
acid.
[0028] We propose and use the name 5-Methylene-Isoleucine for
2-amino-3-methyl-hex-5-enoic acid. Accordingly, the
5-Methytene-Isoleucine side chain has the structure
--CH(CH.sub.3)CH.sub.2CH.dbd.CH.sub.2 which could be
represented
##STR00002##
[0029] The invention also concerns use of 5-Methylene-Isoleucine
for producing Bacitracins and a process for making
5-Methylene-Isoleucine
[0030] 5-Methylene-Isoleucine can be used for production of
Bacitracin J1, Bacitracin J2, Bacitracin J3, Bacitracin K1,
Bacitracin K2, Bacitracin K3 and Bacitracin L.
[0031] The aspects of the invention may be obtained by the features
as set forth in the following description of the invention and/or
the appended patent claims.
DETAILED DESCRIPTION OF THE INVENTION
[0032] Some highly active Bacitracins comprise an uncommon amino
acid side chain which has the following structure:
##STR00003##
[0033] We have proposed and use the names Bacitracin J, K or L for
Bacitracins comprising structure above. The structures of these
Bacitracins are represented in FIG. 1A-G.
Definitions
[0034] "Bacitracins" are peptide compounds comprising the following
structure (with amino acid residue numbering in superscript):
##STR00004##
wherein X is
##STR00005##
and, wherein R is the side chain of the amino acid residue of
Isoleucine, Valine or 5-Methylene-Isoleucine; and wherein Y and Z
are independently the amino acid residue of
[0035] Isoleucine, Valine or 5-Methylene-Isoleucine;
[0036] and wherein
Thz is a Thiazoline ring
##STR00006##
2' coupled to X and 4' coupled to the .alpha.-carbon in Leu; and
wherein Leu is a Leucine amino acid residue Glu is a Glutamine
amino acid residue Lys is a Lysine amino acid residue forming
peptide bonds with Y and Orn while its .epsilon.-amine is coupled
to the .alpha.-carboxyl group of Asparagine by a peptide bond Orn
is an Ornithine amino acid residue Phe is a Phenylalanine amino
acid residue His is a Histidine amino acid residue Asp is an
Aspartic acid amino acid residue Asn is an Asparagine amino acid
residue forming peptide bond with Asp while its .alpha.-carboxyl
group is coupled to the .epsilon.-amine of Lysine by a peptide
bond.
[0037] When used in this application; "Bacitracins" is meant to
embrace any compound having the primary structure above regardless
of the production method. Thus, the term "Bacitracins" includes the
antibiotic compounds naturally produced by Bacillus licheniformis
but also in vitro produced compounds (synthetic) and semisynthetic
compounds having the primary structure above. "Bacitracins" is also
meant to embrace any compound having the primary structure above
regardless of the charge which varies with pH. "Bacitracins" is
also meant to embrace any compound having the primary structure
above regardless of the stereochemistry. "Bacitracins" is also
meant to embrace salts and hydrates of the compounds having the
primary structure above.
[0038] "Bacitracins comprising at least one 5-Methylene-Isoleucine
residue" is meant to embrace any Bacitracin comprising the
structure that would be generated if a isoleucine or Valine
residue(s) was substituted with 5-Methylene-Isoleucine residue(s)
position 1 and/or 5 and/or 8.
[0039] When the N-terminal amino group and/or the Thiazoline ring
of Bacitracins is oxidized, a substantial amount of the
antibacterial activity is lost. For example the low activity
compound Bacitracin F, comprises a keto-thiazole moiety instead of
the amino-thiazoline moiety (J. Org. Chem., vol. 22, 1957, page
1345-1353 by Craig et al).
[0040] "Amino acid" is any compound comprising both an amine and a
carboxyl group. Most proteins are built as a linear polymer of 20
different standard .alpha.-amino acids in L-configuration: Alanine,
Arginine, Asparagine, .Aspartic acid, Cysteine, Glutamine,
Glutamate, Phenylalanine, Glycine Histidine, Isoleucine, Lysine,
Leucine, Methionine, Proline, Serine, Threonine, Tryptophan,
Tyrosine and Valine.
[0041] An "Amino acid residue" is the unit in a peptide which
comprises
--NH--CHR--COOH(C-terminal)
[0042] or
NH.sub.2--CHR--CO (N-terminal)
[0043] or NH--CHR--CO-- (internal) where R is
H Glycine,
CH.sub.3 Alanine,
OH in Serine,
CH.sub.2SH in Cysteine,
CH(CH.sub.3)CH.sub.2CH.sub.3in Isoleucine,
CH.sub.2CH(CH.sub.3).sub.2 in Leucine
CH(CH.sub.3).sub.2 in Valine
[0044] etc.
[0045] An "Amino acid side chain" is the R-group of an "Amino acid
residue". For example, the R-group is
CH(CH)CH.sub.2CH.sub.3in Isoleucine,
CH.sub.2CH(CH.sub.3).sub.2 in Leucine
CH(CH.sub.2).sub.2 in Valine
[0046] "Antibacterial activity" is any activity which
[0047] inhibits the growth, metabolism or reproduction of
bacteria,
[0048] or
[0049] increases the mortality of bacteria.sub.; or
reduces the pathogenicity of bacteria.
[0050] The "positions" of the amino acid residues in Bacitracins
are numbered from the N-terminal which can be Isoleucine, Valine or
5-Methylene-Isoleucine in position 1 (the left end in all figures
showing Bacitracins in this application), Hence, Lys is in position
number 6 and Asn is in position number 12.
[0051] In Bacitracins, the "position 1" is special, because this
amino acid residue is partly incorporated into the Thiazoline ring.
Thus the amino acid residue in position 1 in Bacitracins does not
comprise the usual N-terminal unit:
##STR00007##
but comprises instead:
##STR00008##
coupled to Thiazoline
[0052] A "composition" is any mixture comprising more than two
different compounds, for example a mixture of two active
pharmaceutical ingredients, or a mixture of an active
pharmaceutical ingredient and one or more pharmaceutical
excipients.
[0053] A "pharmaceutical composition" is any composition suitable
for use in vivo. Such compositions can thus be administered
cutaneously, subcutaneously, intravenously, parenterally etc.
[0054] 5-Methylene-Isoleucine comprises two chiral carbon atoms
which independently could be in R or S configuration.
[0055] The Bacitracins comprising at least one
5-Methylene-Isoleucine residue in position 1, 5 or 8 can be used
for inhibiting unwanted bacterial growth both in vitro and in vivo.
These compounds can thus have therapeutic effect if administered to
an animal or a human with a bacterial infection.
[0056] The invention concerns the compound 5-Methylene-Isoleucine
which could be used for production of the new Bacitracins; e.g. by
in vitro synthesis of Bacitracin J1-3, K1-3 or L.
[0057] By substituting Isoleucine or Valine with
5-Methytene-Isoleucine in the methods described in J Org Chem, vol,
61 no. 12, 1996, page 3983-3986 by Lee et at or WO199747313,
Bacitracins with antibacterial activity can be produced.
BRIEF DESCRIPTION OF THE FIGURES
[0058] FIG. 1A shows the structure of Bacitracin with a
5-Methylene-Isoleucine residue in position 5 (=Bacitracin J1)
[0059] FIG. 1B shows the structure of Bacitracin with a
5-Methylene-Isoleucine residue in position 8 (=Bacitracin J2)
[0060] FIG. 1C shows the structure of Bacitracin with a
5-Methylene-Isoleucine residue in position 1 (=Bacitracin J3)
[0061] FIG. 1D shows the structure of Bacitracin with
5-Methylene-Isoleucine residues in position 5 and 8 (=Bacitracin
K1)
[0062] FIG. 1E shows the structure of Bacitracin with
5-Methylene-Isoleucine residues in position 1 and 5 (=Bacitracin
K2)
[0063] FIG. 1F shows the structure of Bacitracin with
5-Methylene-Isoleucine residues in position 1 and 8 (=Bacitracin
K3)
[0064] FIG. 1G shows the structure of Bacitracin with
5-Methylene-Isoleucine residues in position 1, 5 and 8 (=Bacitracin
L)
[0065] FIG. 2 shows a route tier production of
5-Methylene-Isoleucine as disclosed in the examples
[0066] The invention is defined by the claims and not by the
following illustrative examples:
EXAMPLES
Example 1
(2S,3R)-2-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-e-
noic acid methyl ester and
(2S,3S)-2-(1,3-dioxo-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-enoic
acid methyl ester
[0067] Vinylmagnesium bromide in THE (1.0 M, 83.64 mL, 83.64 mmol)
is added to a precooled (-10.degree. C.) mixture of (2S,3S) and
(2S,3R)-4-Iodo-N-phataloylvaline methyl ester (26.96 g, 69.68
mmol), CuCl.sub.2(0.47 g, 3.5 mmol), dry LiCl (0.29 g, 7.0 mmol) in
THF (100 mL). The reaction mixture is stirred over night at this
temperature. The reaction mixture is added to saturated ammonium
chloride solution (aq) (300 mL). The phases are separated and the
aqueous phase is extracted with diethyl ether (3.times.100 mL). The
combined organic phases are washed with saturated brine (100 mL),
dried (MgSO.sub.4) and the solvents are removed at reduced
pressure. 10 g of the product mixture is isolated The (2S,3S) and
(2S,3R) products are separated by column chromatography.
Example 2 (2S,3R)-2-Amino-3-methyl-hex-5-enoic acid
[0068] (2S,3R)-2-(1,3-Dioxo-1,3-dihydro-isoindol
-2-yl)-3-methyl-hex-5-enoic acid methyl ester (2.01 g, 7.00 mmol)
is dissolved in a 2:1 mixture of 6 N hydrochloric acid and glacial
acetic acid (62.5 mL), and the solution is heated at reflux for 4
h. The solution is cooled to room temperature and concentrated
under reduced pressure.
[0069] The product is taken up in water and the solution is
filtered. The filtrate is concentrated under reduced pressure and
the residue is dissolved in water, then the solution is applied to
a column of amberlite IR 120 cation exchange resin (NH.sub.4.sup.+
form). The column is washed with water (2.5 L), then eluted with
aqueous ammonia solution (2.5 L). The eluate is boiled until no
ammonia can be detected, then concentrated under reduced pressure
affording the title compound (0.5 g, 3.50 mmol).
Example 3 (2S,3S)-2-Amino-3-methyl-hex-5-enoic acid
[0070]
(2S,3S)-2-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-enoi-
c acid methyl ester (2.01 g, 7.00 mmol) is dissolved in a 2:1
mixture of 6 N hydrochloric acid and glacial acetic acid (62.5 mL),
and the solution is heated at reflux for 4 h. The solution is
cooled to room temperature and concentrated under reduced
pressure.
[0071] The product is taken up in water and the solution is
filtered. The filtrate is concentrated under reduced pressure and
the residue is dissolved in water, then the solution is applied to
a column of amberlite IR 120 cation exchange resin (NH.sub.4.sup.+
form). The column is washed with water (2.5 L), then eluted with
aqueous ammonia solution (2.5 L). The eluate is boiled until no
ammonia can be detected, then concentrated under reduced pressure
affording the title compound (0.5 g, 150 mmol).
Example 4
(2R,3R)-2-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-e-
noic acid methyl ester and (2R,3S)-2-(1,3-dioxo
-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-enoic acid methyl
ester
[0072] Vinylmagnesium bromide in THF (1.0 M, 83.64 mL, 83.64 mmol)
is added to a precooled (-10.degree. C.) mixture of (2S,3S) and
(2S,3R)-4-Iodo-N-phataloyivaline methyl ester (26.96 g, 69.68
mmol), CuCl.sub.2 (0.47 g, 3.5 mmol), dry LiCl (0.29 g, 7.0 mmol)
in THF (100 mL). The reaction mixture is stirred over night at this
temperature. The reaction mixture is added to saturated ammonium
chloride solution (aq) (300 mL). The phases are separated and the
aqueous phase is extracted with diethyl ether (3.times.100 mL). The
combined organic phases are washed with saturated brine (100 mL),
dried (MgSO.sub.4) and the solvents are removed at reduced
pressure. 10 g of the product mixture is isolated. The (2R,3S) and
(2R,3R) products are separated by column chromatography.
Example 5 (2R,3R)-2-Amino-3-methyl-hex-5-enoic acid
[0073]
(2R,3R)-2-(1,3-Dioxo-1,3-dihydro-isoindol-2-yl)-3-methyl-hex-5-enoi-
c acid methyl ester (2.01 g, 7.00 mmol) is dissolved in a 2:1
mixture of 6 N hydrochloric acid and glacial acetic acid (62.5 mL),
and the solution is heated at reflux for 4 h. The solution is
cooled to room temperature and concentrated under reduced
pressure.
[0074] The product is taken up in water and the solution is
filtered. The filtrate is concentrated under reduced pressure and
the residue is dissolved in water, then the solution is applied to
a column of amberlite IR 120 cation exchange resin (NH.sub.4.sup.+
form). The column is washed with water (2.5 then eluted with
aqueous ammonia solution (2.5 L). The eluate is boiled until no
ammonia can be detected, then concentrated under reduced pressure
affording the title compound (0.5 g, 3.50 mmol).
Example 6 (2R,3S)-2-Amino-3-methyl-hex-5-enoic acid
[0075] (2R,3S)-1,3-Dioxo-1,3-dihydro-hex-5-enoic acid methyl ester
(2.01 g, 7.00 mmol ) is dissolved in a 2:1 mixture of 6 N
hydrochloric acid and glacial acetic acid (62.5 mL), and the
solution is heated at reflux for 4 h. The solution is cooled to
room temperature and concentrated under reduced pressure. The
product is taken up in water and the solution is filtered. The
filtrate is concentrated under reduced pressure and the residue is
dissolved in water, then the solution is applied to a column of
amberlite IR 120 cation exchange resin (NH.sub.4.sup.+ form). The
column is washed with water (2.5 L), then eluted with aqueous
ammonia solution (2.5 L). The eluate is boiled until no ammonia can
be detected, then concentrated under reduced pressure affording the
title compound (0.5 g, 3.50 mmol).
* * * * *