U.S. patent number RE40,480 [Application Number 11/405,041] was granted by the patent office on 2008-09-02 for 1,3-propane diol esters and ethers and methods for their use in drug delivery.
This patent grant is currently assigned to Scarista Limited. Invention is credited to Paul Bradley, Sherri Clarkson, David B. Horrobin, Philip Knowles, Mehar Manku, Austin McMordie, Andrea Pitt, Peter Redden, Paul Wakefield.
United States Patent |
RE40,480 |
Clarkson , et al. |
September 2, 2008 |
1,3-Propane diol esters and ethers and methods for their use in
drug delivery
Abstract
The invention relates to compounds of formula: ##STR00001##
wherein R.sup.1 is selected from the group consisting of fatty acid
acyl groups of 12 to 30 carbon atoms and fatty alcohol groups of 12
to 30 carbon atoms, and wherein R.sup.2 is selected from the group
consisting of H, fatty acid acyl of 12 to 30 carbon atoms and fatty
alcohol groups of 12 to 30 carbon atoms, the same as or different
from R.sup.1, and the residue of a nutrient, drug, or other
bioactive compound, and to the use of these compounds to deliver
drugs and other bioactive compounds.
Inventors: |
Clarkson; Sherri (Douglas,
GB), Manku; Mehar (Hollywood, GB), Knowles;
Philip (Cumbria, GB), Redden; Peter (Oakville,
CA), Pitt; Andrea (Barnard Castle, GB),
Bradley; Paul (Hathern, GB), Wakefield; Paul
(Cumbria, GB), McMordie; Austin (Craigavon,
GB), Horrobin; David B. (Guildford, GB) |
Assignee: |
Scarista Limited (Douglas, Isle
of Man, GB)
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Family
ID: |
39718531 |
Appl.
No.: |
11/405,041 |
Filed: |
May 1, 1996 |
PCT
Filed: |
May 01, 1996 |
PCT No.: |
PCT/GB96/01053 |
371(c)(1),(2),(4) Date: |
January 28, 1998 |
PCT
Pub. No.: |
WO96/34846 |
PCT
Pub. Date: |
November 07, 1996 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
Issue Date |
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11119495 |
Apr 29, 2005 |
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Reissue of: |
08945667 |
Jan 28, 1998 |
06555700 |
Apr 29, 2003 |
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Foreign Application Priority Data
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May 1, 1995 [GB] |
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9508823 |
Aug 21, 1995 [GB] |
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9517107 |
Mar 15, 1996 [GB] |
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9605440 |
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Current U.S.
Class: |
514/29; 548/496;
549/39; 554/82; 560/38; 560/128; 554/227; 514/420 |
Current CPC
Class: |
Y02A
50/30 (20180101); Y02A 50/411 (20180101) |
Current International
Class: |
C07C
57/13 (20060101) |
Field of
Search: |
;548/496 ;549/39
;560/38,128 |
References Cited
[Referenced By]
U.S. Patent Documents
Foreign Patent Documents
|
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|
|
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0 018 342 |
|
Feb 1983 |
|
EP |
|
0 087 864 |
|
Sep 1983 |
|
EP |
|
0 139 480 |
|
May 1985 |
|
EP |
|
0 139 780 |
|
May 1985 |
|
EP |
|
0161114 |
|
Nov 1985 |
|
EP |
|
0 173 478 |
|
Mar 1986 |
|
EP |
|
0 057 797 |
|
Jun 1986 |
|
EP |
|
0 184 058 |
|
Jun 1986 |
|
EP |
|
0 056 189 |
|
Aug 1986 |
|
EP |
|
0 222 155 |
|
May 1987 |
|
EP |
|
0 246 540 |
|
Nov 1987 |
|
EP |
|
0 161 422 |
|
Mar 1989 |
|
EP |
|
0319126 |
|
Jun 1989 |
|
EP |
|
0 321 128 |
|
Jun 1989 |
|
EP |
|
0 393 920 |
|
Oct 1990 |
|
EP |
|
0 405 873 |
|
Jan 1991 |
|
EP |
|
0 405 874 |
|
Jan 1991 |
|
EP |
|
0 574 312 |
|
Dec 1993 |
|
EP |
|
0 611 569 |
|
Aug 1994 |
|
EP |
|
0 675 103 |
|
Oct 1995 |
|
EP |
|
1 782 |
|
Apr 1963 |
|
FR |
|
0 888 162 |
|
Jan 1962 |
|
GB |
|
1 135 647 |
|
Dec 1968 |
|
GB |
|
1 293 277 |
|
Oct 1972 |
|
GB |
|
1 493 098 |
|
Nov 1977 |
|
GB |
|
1 529 762 |
|
Oct 1978 |
|
GB |
|
1 556 197 |
|
Nov 1979 |
|
GB |
|
2 161 477 |
|
Sep 1987 |
|
GB |
|
57-067511 |
|
Apr 1982 |
|
JP |
|
61-129190 |
|
Jun 1986 |
|
JP |
|
2-129119 |
|
May 1990 |
|
JP |
|
4-99784 |
|
Mar 1992 |
|
JP |
|
05-051355 |
|
Mar 1993 |
|
JP |
|
WO 91/09831 |
|
Jul 1991 |
|
WO |
|
WO 94/26262 |
|
Nov 1994 |
|
WO |
|
WO 95/04030 |
|
Feb 1995 |
|
WO |
|
WO 96/33155 |
|
Oct 1996 |
|
WO |
|
Other References
Breusch et al., Chemische Berichte, vol. 88, pp. 1511-1519, 1955.
cited by examiner .
Langen et al., Chemical Abstracts, vol. 92, No. 1, #461a, 1980.
cited by examiner .
Lion Corp., Patent Abstracts of Japan, vol. 6, No. 143(C-117),
1982. cited by examiner .
Vajdi et al., Chemical Abstracts, vol. 95, No. 5, #41676e, 1981.
cited by examiner .
Hounsom et al., Oxidative Stress and Disease (2000), 4(Antioxidants
in Diabetes Management), 155-172. cited by examiner .
Watanabe et al, Chemical Abstracts, vol. 107, No. 3, #23659p, 1987.
cited by examiner .
Bertelsen, O. et al., "Structural Elucidation of Alkyl-Branched
Chain Aliphatic Alcohols and Fatty Acids by Mass Spectrometry of
their Respective Alkyl Nicotinate and Picolinylcarboxylate
Derivatives," Chemical Abstracts, 104:108841 (1986). cited by
examiner .
Christie, W.W., et al., "Mass Spectra of the Picolinyl Ester
Derivatives of Some Isomeric Dimethylene-Interrupted
Octadecadiynoic Acids," Chemical Abstracts, 109:210389(1988). cited
by examiner .
Christie, W.W., et al., "Mass Spectra of the Picolinyl Esters of
Isomeric Mono-and Dienoic Fatty Acids," Chemical Abstracts
107:55235 (1987). cited by examiner .
Deterding, L. et al., "Fast-Atom-Bombardment and Tandem Mass
Spectrometry for Determining Structures of Fatty Acids and Their
Picolyl Ester Derivatives," Chemical Abstracts, VI. 110:38801
(1989). cited by examiner .
Deterding, L., et al., "Tandem Mass Spectrometry for Identifying
Fatty Acid Derivatives that Undergo Charge-Remote Fragmentations,"
Chemical Abstracts 110:56917 (1989). cited by examiner .
Harvey, D.J., Picolinyl Derivatives for the structural
Determination of Fatty Acids by Chemical Abstracts 101:166564
(1984). cited by examiner .
Harvey, D.J., "Pyridine-Containing Derivatives for the Structural
Elucidationof the Alkyl Chains of Lipids by Mass Spectrometry and a
Comparison with the Spectra of Related Heterocyclic Derivatives,"
Chemical Abstracts, 114:206360 (1991). cited by examiner .
Jie, et al., "Mass Spectra of Picolinyl Ester Derivatives of Some
Conjugated Diacetylenic Fatty Acids," Chemical Abstracts,
118:191384 (1993). cited by examiner .
Kimura, K. et al., Higher Unsaturated Fatty Alcohol Esters Having
Antiucler Activity, Chemical Abstracts, vol. 87, 1997 Abstract No.
53462e. cited by examiner .
Koori, S., "Unsaturated Higher Akuogatuc Esters of Nicotinic Acid,"
Chemical Abstracts, vol. 77, Abstract No. 164488f (1972). cited by
examiner .
Kumokawa, Y. et al. .gamma.-Linolenic Acid Derivatives As Platelet
Aggregation Inhibitors, Chemical Abstracts, vol. 105, 1986,
Abstract No. 78532f. cited by examiner .
Spitzer, V. et al., "Curupira Tefeensis II: Occurrence of
Acetylenic Fatty Acids," Fat Sci. Technol., No. 5, (1991) pp.
169-174. cited by examiner .
Terumo Corp., "Trienoic Fatty Acid Pyridylmethyl Esters," Chemical
Abstracts, vol. 101, 1984 Abstract No. 171104v. cited by
examiner.
|
Primary Examiner: Anderson; Rebecca L
Attorney, Agent or Firm: Buchanan Ingersoll & Rooney
PC
Parent Case Text
.Iadd.Notice: More than one reissue application has been filed for
reissue on U.S. Pat. No. 6,555,700. U.S. application No. 11/405,041
(the present application) filed Apr. 14, 2006 is a divisional
reissue of U.S. application No. 11/119,495 filed Apr. 29, 2005
which is a reissue of U.S. application No. 08/945,667 filed Jan.
28, 1998, now U.S. Pat. No. 6,555,700 which is a 371 of
PCT/GB96/01053 filed May 1, 1996..Iaddend.
This application is a 371 of PCT/GB96/01053, filed May 1, 1996.
Claims
What is claimed is:
.[.1. A compound having the formula:.]. ##STR00004## .[.wherein
R.sup.1 is selected from the group consisting of fatty acid acyl
groups of 12 to 30 carbon atoms and fatty alcohol groups of 12 to
30 carbon atoms, and wherein R.sup.2 is selected from the group
consisting of fatty acid acyl groups of 12 to 30 carbon atoms and
fatty alcohol groups of 12 to 30 carbon atoms, the same as or
different from R.sup.1, the fatty acid acyl or alcohol groups
R.sup.2 being selected from the group consisting of
.gamma.-linoleic acid (GLA), dihomo-.gamma.-linolenic acid (DGLA),
arachidonic acid (AA), adrenic acid, stearidonic acid (SA),
eicosapentaenoic acid (EPA), docosapentaenoic acid n-3,
docosahexaenoic acid (DHA), columbinic acid (CA), parinaric acid
and conjugated linoleic acid (cLA) groups..].
.[.2. The compound according to claim 1, wherein each said fatty
acid or fatty alcohol group has 16 to 30 carbon atoms..].
.[.3. The compound according to claim 1, having a phosphate,
succinate, or other difunctional-acid linking moiety between
R.sup.1 and the corresponding diol oxygen, between R.sup.2 and the
corresponding diol oxygen, or both..].
.[.4. The compound according to claim 1, wherein one of R.sup.1 and
R.sup.2 is an acyl moiety corresponding to an acid selected from
the group consisting of .gamma.-linolenic acid (GLA) and
dihomo-.gamma.-linolenic acid (DGLA), and the other of R.sup.1 and
R.sup.2 is an acyl moiety corresponding to an acid selected from
the group consisting of .gamma.-linolenic acid (GLA),
dihomo-.gamma.-linolenic acid (DGLA), stearidonic acid (SA),
eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), conjugated
linoleic acid (cLA), and columbinic acid (CA)..].
.[.5. The compound according to claim 1, wherein one of R.sup.1 and
R.sup.2 is an acyl moiety corresponding to an acid selected from
the group consisting of arachidonic acid (AA), and the other is an
acyl moiety corresponding to an acid selected from the group
consisting of arachidonic acid (AA), .gamma.-linolenic acid (GLA),
dihomo-.gamma.-linolenic acid (DGLA), docosahexaenoic acid (DHA),
and eicosapentaenoic acid (EPA)..].
.[.6. The compound according to claim 1, wherein one of the R.sup.1
and R.sup.2 is an acyl moiety corresponding to eicosapentaenoic
acid (EPA) and the other is an acyl moiety corresponding to an acid
selected from the group consisting of eicosapentaenoic acid (EPA)
and docosahexaenoic acid (DHA)..].
.[.7. A pharmaceutical composition comprising an effective amount
of the compound of claim 1 and a suitable diluent or
carrier..].
.[.8. A compound according to claim 1 having the following
1,3-propane diol linked structure:.]. ##STR00005## .[.wherein
R.sup.1 and R.sup.2 are pairs of fatty acids selected from the
group of pairs of fatty acids consisting of: .gamma.-linolenic acid
and oleic acid; .gamma.-linolenic acid and .gamma.-linolenic acid;
eicosapentaenoic acid and eicosapentaenoic acid; .gamma.-linolenic
acid and eicosapentaenoic acid; .gamma.-linolenic acid and
docosahexaenoic acid; arachidonic acid and docosahexaenoic acid;
arachidonic acid and eicosapentaenoic acid; .gamma.-linolenic acid
and arachidonic acid; .gamma.-linolenic acid and stearidonic acid;
stearidonic acid and docosahexaenoic acid; arachidonic acid and
stearidonic acid; dihomo-.gamma.-linolenic acid and
dihomo-.gamma.-linolenic acid; dihomo-.gamma.-linolenic acid and
.gamma.-linolenic acid; dihomo-.gamma.-linolenic acid and
stearidonic acid; dihomo-.gamma.-linolenic acid and arachidonic
acid; dihomo-.gamma.-linolenic acid and eicosapentaenoic acid;
dihomo-.gamma.-linolenic acid and docosahexaenoic acid; arachidonic
acid and arachidonic acid; eicosapentaenoic acid and stearidonic
acid; eicosapentaenoic acid and docosahexaenoic acid;
docosahexaenoic acid and docosahexaenoic acid; conjugated linoleic
acid and conjugated inoleic acid, conjugated linoleic acid and
.gamma.-linolenic acid; conjugated linoleic acid and
dihomo-.gamma.-linolenic acid; conjugated linoleic acid and
arachidonic acid; conjugated hinoleic acid and stearidonic acid;
conjugated linoleic acid and eicosapentaenoic acid; conjugated
linoleic acid and docosahexaenoic acid; columbinic acid and
columbinic acid; columbinic acid and .gamma.-linolenic acid;
columbinic acid and dihomo-.gamma.-linolenic acid; columbinic acid
and arachidonic acid; columbinic acid and stearidonic acid;
columbinic acid and eicosapentaenoic acid; and columbinic acid and
docosahexaenoic acid..].
.[.9. The compound according to claim 8, wherein R.sup.1 and
R.sup.2 are both docosahexaenoic acid moieties..].
.[.10. The compound according to claim 8, wherein R.sup.1 and
R.sup.2 are both eicosapentaenoic acid moieties..].
.[.11. A pharmaceutical composition comprising an effective amount
of the compound of 8, 9 or 10 and a suitable diluent or
carrier..].
.Iadd.12. A compound having the formula:
R.sup.1OCH.sub.2--CH.sub.2--CH.sub.2OR.sup.2 wherein R.sup.1 is
selected from the group consisting of fatty acid acyl groups of 12
to 30 carbon atoms and fatty alcohol groups of 12 to 30 carbon
atoms, and wherein R.sup.2 is lipoic acid..Iaddend.
.Iadd.13. A compound having the formula:
R.sup.1OCH.sub.2--CH.sub.2--CH.sub.2OR.sup.2 wherein R.sup.1 is
selected from the group consisting of fatty acid acyl groups of 12
to 30 carbon atoms and fatty alcohol groups of 12 to 30 carbon
atoms, and wherein R.sup.2 is a tryptophan, phenylalanine, GABA,
fluoextine, or tranylcypromine moiety..Iaddend.
.Iadd.14. The compound according to claim 13, wherein R.sup.1 is is
.gamma.-linolenic acid (GLA) and R.sup.2 is
tryptophan..Iaddend.
.Iadd.15. The compound according to claim 13, wherein R.sup.1 is is
.gamma.-linolenic acid (GLA) and R.sup.2 is
phenylalanine..Iaddend.
.Iadd.16. The compound according to claim 13, wherein R.sup.1 is is
.gamma.-linolenic acid (GLA) and R.sup.2 is GABA..Iaddend.
.Iadd.17. The compound according to claim 15, wherein R.sup.1 is is
.gamma.-linolenic acid (GLA) and R.sup.2 is
phenylalanine..Iaddend.
.Iadd.18. The compound according to claim 15, wherein R.sup.1 is is
.gamma.-linolenic acid (GLA) and R.sup.2 is GABA..Iaddend.
Description
FIELD
The specification relates to the presentation of bioactives, in
which term we include a drug, essential nutrient or any other
compound to be administered to the human or animal body in therapy
or maintenance of health.
In particular, the specification relates to the presentation of
such bioactives in a form in which they are lipophilic so that they
can pass lipid barriers in the body readily, or to the presentation
of two bioactives in the same molecule (where at least one of the
bioactives is a fatty acid or fatty alcohol), or to the
presentation of bioactives in a form which serves both aims and/or
the aims of ready synthesis of such compounds without a chiral
centre. From a drug regulatory viewpoint it is a great advantage to
have two bioactives presented as a single molecule rather than as
two separate entities. There may also be advantages in presenting
known bioactives in novel ways. Those advantages include increased
lipophilicity, the additives effects of two bioactives which are
not normally presented together, and the sometimes synergistic
effects of such bioactives.
The invention concerns the linking of bioactives through certain
link molecules, considered in detail later herein, and the
synthesis of a range of compounds some of which are entirely novel
in themselves, while others are novel in the sense of their
usefulness in therapy and/or the maintenance of health. Discussion
is however, also given of compounds using other link molecules not
currently claimed, and of directly linked bioactives, disclosed for
example in EPA-0 393 920 concerning fatty acids and antivirals, and
in co-pending EP-95301315.8 (published as EPA-0 675 103) concerning
fatty acids and non-steroidal anti-inflammatory drugs.
Published Material
Concepts such as are discussed above have received no great
attention in the published patent and general literature but there
is material on certain specific natural diol derivatives and on
nutritional and pharmaceutical uses of certain specific diol
esters. A source paper in the general literature is Bergelson et al
(Biochim. Biophys. Acta 1 16 (1966) 511-520) describing inter alia
long chain diesters of 1,3-propane diol. Little is said of the acid
moieties but dioleates are identified. In the patent literature
edible fat mimetics are for example proposed by Nabisco in EPA-0
405 873 and EPA-0 405 874 and include linolenic acid esters (this
term indicating the "alpha" isomer when not qualified otherwise)
and arachidonic acid esters of, apparently, 1,4-butane diol.
Unilever's U.K. specification 2 161 477 (equivalent to EPA-0 161
114) concerns the growth and economic yield of plants, using inter
alia 1,3-propane diol esters of linoleic acid and linolenic acid
(again no doubt the alpha isomer). Anti-ulcer drugs of
2,3-butanediol esters are described in SS Pharmaceutical Co's EPA-0
056 189. Sundry pharmaceutical actions of propane-1,3-diol esters
of short chain fatty acids are disclosed in Sanofi EPA-0 018 342.
More distantly perhaps, Terumo K. K. in EPA-0 222 155 links
5-fluoro uracil to alpha linolenic acid, dihomo gamma linolenic
acid, or eicosapentaenoic acid through a group --CH(R)--O-- where
R=methyl etc as, inter alia, anti-cancer agents.
Lipid Barriers
Many drugs act at the cell membrane surface by combining with cell
surface receptors, or alternatively are taken into cells by
specific transport systems. However, there are many drugs which,
while they act within cells by modifying one of many different
functions such as nucleic acid functions, the actions of
intracellular enzymes, or the behaviour of systems like the
lysosomes or the microtubules, are not able to penetrate cells
effectively. There may be no receptors and transport systems with
which they can link, or these systems may transport the drug into
the cell at a less then optimum rate. Equally drugs may penetrate
intracellular membranes such as mitochondrial and nuclear membranes
at less than optimum rates.
There are other barriers to drug movements which are recognised as
important. One of particular significance is the blood-brain
barrier, which has many of the characteristics of the cell
membrane. There are many drugs which have difficulty in reaching
adequate concentrations in the brain because of this barrier.
Another is the skin: until a few years ago drugs were applied to
the skin only if their purpose was to act on the skin. However, it
has been recognised that the skin can be an appropriate route for
getting drugs with systemic actions into the body, and as a result
more and more compounds are being administered by variations of
patch technology.
All three types of barriers, the cell membrane and intra-cellular
membranes, the blood-brain barrier and the skin have an important
feature in common, they are substantially composed of lipids. What
this means is that they are impermeable to primarily water-soluble
drugs unless these drugs can be carried across the membrane by a
receptor or transport system. In contrast, lipophilic substances
are able to cross the barriers more readily without the need for
any specific receptor or transport system.
Classes of Bioactives Requiring Passage Through Lipid Barriers
Drugs whose pharmacokinetic behaviour may be improved by increased
lipophilicity, listed by route of entry, are as follows: Cell
entry: drugs particularly likely to benefit are those that act
primarily intracellularly. These include: a. All anti-inflammatory
drugs, whether steroid or non-steroid b. All cytotoxic drugs used
in the management of cancer; c. All antiviral drugs; d. All other
drugs that have to enter cells in order to achieve optimum effects,
in particular drugs which act on DNA or RNA, or on enzymes located
intracellularly, or on second messenger systems, or on
microtubules, mitochondria, lysosomes, or any other intracellular
organelle. e. Steroid hormones and other hormones that act
intracellularly, such as oestrogens, progestins, androgenic
hormones and dehydroepiandrosterone. 2. Blood-brain barrier: all
drugs acting on the central nervous systems will have their
transport improved by this technique. This includes all drugs used
in psychiatry, all drugs used in cerebral infections with any
organism or in cerebral cancer and all other drugs acting on nerve
cells such as anti-epileptic drugs and others acting on
neurological disorders such as multiple sclerosis, amyotrophic
lateral sclerosis, Huntington's chorea and others. 3. Skin: as with
the blood-brain barrier, all drugs that may be required to
penetrate the skin to achieve a systemic effect will benefit from
their conversion to a fatty acid derivatives.
For example, the approach discussed is applicable to amino acids.
Of particular interest are those which seem to play roles in the
regulation of cell function as well as acting as components of
proteins. Examples include tryptophan (a precursor of
5-hydroxytryptamine [5-HT], a key regular of nerve and muscle
function), phenylalanine (a precursor of catecholamines) and
arginine (a regulator of the synthesis of nitric oxide which also
plays important roles in controlling cellular activities).
Properties Conferred Generally
Generally the compounds proposed herein have many advantages in
addition to their lipophilicity. Two moieties of a given fatty acid
or even a single moiety may be delivered, in a form which is
readily incorporated into the body as an oral, parenteral or
topical formation; which is very well tolerated with none of the
side effects associated, for example, with free fatty acids; which
is not too stable to be properly utilised; which need have no
chiral centre; and which is much more readily synthesised than the
corresponding triglyceride with three moieties of the same fatty
acid attached. Whereas triglycerides are well tolerated and well
utilised, they are less desirable than the proposed compounds
because they are more difficult to synthesise and may have a chiral
centre with multiple potential isomers. Moreover with triglycerides
the fatty acids may relatively easily migrate from one position to
another creating new molecules not present in the original
preparation. This obviously causes problems, particularly in the
context of drug regulation where such instability may be
unacceptable.
When two different fatty acids are to be delivered, the advantages
are as before plus the ability to administer simultaneously two
materials with different biological actions in a single molecule.
This avoids the regulatory problems which ensue when two materials
are administered as separate compounds, as well as the issues which
arise where there is the possibility of chiral centres. When two
drugs are delivered as separate molecules, regulatory authorities
normally require each drug to be studied alone as well as in
combination. If the two are combined in a single molecule, only the
single molecule needs to be studied, greatly reducing the cost of
development.
Where actives other than fatty acids are present there are similar
advantages. The compounds allow drugs or other compounds to be
administered in the form of relatively-lipophilic compounds which
are non-chiral (unless the drugs or other compounds are themselves
chiral), which release the active moieties relatively easily, and
which are well tolerated on oral, topical or parenteral
administration. Their lipophilicity enables them to be absorbed
partially through the lymphatic system, so by-passing the liver; to
cause less gastrointestinal irritation than with many compounds;
and to facilitate transport of drugs and other agents across
lipophilic barriers such as the skin, the cell membrane and the
blood-brain barrier.
There is evidence that interesting specific properties in addition
to ready passage of lipid barriers can be conferred on many drugs
by making them more lipophilic. These properties include prolonged
duration of action, reduction of side effects especially
gastrointestinal, bypassing of first-pass liver metabolism and,
potentially, site specific delivery of different materials.
Fatty Acid Derivatives; Effects of the Fatty Acids
The transport of actives across lipid membranes may be improved by
linking them directly or via intermediate links to, in particular,
gamma-linolenic acid (GLA) or dihomo-gamma-linolenic acid (DGLA),
two fatty acids which in themselves have a range of desirable
effects. These links also enable bioactive substances to be
co-delivered in the same molecule with fatty acids which in
themselves have desirable actions, irrespective of any transport
advantages. Other fatty acids, such as any of the essential fatty
acids (EFAs) and in particular the twelve natural acids of the n-6
and n-3 series EFAs (FIG. 1), can be used. Of these twelve,
arachidonic acid, adrenic acid, stearidonic acid, eicosapentaenoic
acid and docosahexaenoic acid are of particular interest because
they in themselves have particularly desirable effects.
Furthermore, any fatty acid, suitably C.sub.12-C.sub.30 or
C.sub.16-C.sub.30 and desirably with two or more cis or trans
carbon-carbon double bonds may also be of use. Use may be in the
form of the fatty acid or the corresponding fatty alcohol.
Conjugated linoleic and columbinic acids are examples of fatty
acids which in themselves have valuable properties and are likely
to be of particular use: References to fatty acids are accordingly
to be read herein as to both forms, except where the chemistry of
one or the other specifically is under discussion. The desirable
properties of GLA and DGLA however, make them especially valuable
for the purpose.
The essential fatty acids, which in nature are of the all--cis
configuration, are systematically named as derivatives of the
corresponding octadecanoic, eicosanoic or docosanoic acids, e.g.
z,z-octadeca-9,12-dienoic acid or
z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoic acid, but numerical
designations based on the number of carbon atoms, the number of
centres of unsaturation and the number of carbon atoms from the end
of the chain to where the unsaturation begins, such as,
correspondingly, 18:2n-6 or 22:6n-3 are convenient. Initials, e.g.,
EPA and shortened forms of the name e.g. eicosapentaenoic acid are
used as trivial names in some of the cases.
TABLE-US-00001 FIG. 1 n-6 EFA's n-3 EFA's 18:2n-6 18:3n-3 (Linoleic
acid, LA) (.alpha.-Linolenic acid, ALA) .dwnarw.
.delta.-6-desaturase .dwnarw. 18:3n-6 18:4n-3 (.gamma.-Linolenic
acid, GLA) (Stearidonic acid, SA) .dwnarw. elongation .dwnarw.
20:3n-6 20:4n-3 (Dihomo-.gamma.-linolenic acid, DGLA) .dwnarw.
.delta.-5-desaturase .dwnarw. 20:4n-6 20:5n-6 (Arachidonic acid,
(Eicosapentaenoic acid, AA) EPA) .dwnarw. elongation .dwnarw.
22:4n-6 22:5n-3 (Adrenic acid) .dwnarw. .delta.-4-desaturase
.dwnarw. 22:5n-6 22:6n-3 (Docosahexaenoic acid, DHA)
GLA and DGLA
In their own right GLA and DGLA have been shown to have
anti-inflammatory effects, to lower blood pressure, to inhibit
platelet aggregation, to lower cholesterol levels, to inhibit
cancer cell growth, to reduce dyskinetic movements, to relieve
breast pain, to improve calcium absorption and enhance its
deposition in bone, to reduce the adverse effects of ionising
radiation, to treat various psychiatric disorders, to cause
vasodilation, to improve renal function, to treat the complications
of diabetes, to dilate blood vessels and so on. Activities linked
to GLA and DGLA will therefore not only become more lipophilic,
enhancing penetration across all membranes, the skin and the blood
brain barrier, but are also likely to exhibit new and additional
therapeutic effects. The fatty acid compounds may thus be mutual
bipartate prodrugs (if linked directly) or mutual tripartate
prodrugs (if connected via a link).
Other fatty acids likely to be of especial value in this context
are arachidonic acid and docosahexaenoic acid which are major
constituents of all cell membranes; adrenic acid; and stearidonic
acid and eicosapentaenoic acid which have ranges of desirable
properties similar to those of GLA and DGLA. Fatty acids not
included in the fatty acids of FIG. 1 which are of particular
interest are conjugated linoleic acid (cLA) and columbinic acid
(CA). cLA has a range of interesting effects in treating and
preventing cancer, in promoting growth particularly of
protein-containing tissues, in preventing and treating
cardiovascular disease and as an antioxidant. CA has many of the
properties of essential fatty acids.
Classes of Actives Having Mutual Efficacy with Bioactive Fatty
Acids
Kinds of actives to be incorporated in compounds as set out herein
may be broadly stated: a) Drugs including antibiotics,
antiprotozoals, antipsychotics, antidepressants and NSAIDS and
compounds used in the treatment of cardiovascular, respiratory,
dermatological, psychiatric, neurological, renal, muscular,
gastrointestinal, reproductive and other diseases and in cancer. b)
Hormones c) Amino acids d) Vitamins particularly of the B group,
and other essential nutrients. e) Cytokines and peptides f)
Neurotransmitters and neurotransmitter precursors. g) Phospholipid
head groups such as inositol, choline, serine and ethanolamine,
which may be linked directly or via the phosphate moiety. h)
Aromatic fatty acids such as phenylacetic acid, phenyl butyric acid
and cinnamic acid which are of particular value in cancer
treatment.
Efficacy
The combination of the therapeutic effect of a drug with the
therapeutic effect of a fatty acid may be considered through
examples: a) Psychotropic drugs may be linked to fatty acids such
as GLA, DGLA, arachidonic acid, eicosapentaenoic acid or
docosahexaenoic acid which have important roles in brain function,
so providing a dual therapeutic effect. b) Drugs used for the
treatment of cardiovascular disease may be attached to a fatty acid
which also has value in such treatment, such as eicosapentaenoic
acid which lowers triglyceride levels and inhibits platelet
aggregation, or GLA or DGLA which lower cholesterol levels and have
vasodilator action, or arachidonic acid which is a potent
cholesterol lower agenting, or DHA which has anti-arrhythmic
properties. c) Drugs used in the treatment of any form of
inflammation may be linked to a fatty acid such as gammalinolenic
acid, dihomo-gammalinolenic acid or eicosapentaenoic acid or
docosahexaenoic acid which also has anti-inflammatory action. d)
Drugs used in the management of osteoporosis may be linked to GLA
or DGLA which enhance the incorporation of calcium into bone, or to
EPA or DHA which reduces urinary calcium excretion. e) Drugs used
in skin disease may be linked to GLA or DGLA which have
anti-inflammatory effects on the skin. f) Drugs used in cancer may
be linked to GLA, DGLA, arachidonic acid, EPA or DHA which have
anticancer effects in their own right and which may reverse
resistance to anticancer drugs.
Concepts Applied to Essential Fatty Acids as Bioactives
The essential fatty acids (EFAs) as already referred to, and well
known, consist of a series of twelve compounds. Although linoleic
acid, the parent compound of the n-6 series, and alpha-linolenic
acid, the parent compound of the n-3 series, are the main dietary
EFAs, these substances as such have relatively minor roles in the
body. In order to be fully useful to the body, the parent compounds
must be metabolised to longer chain and more highly unsaturated
compounds. In quantitative terms, as judged by their levels in cell
membranes and in other lipid reactions dihomogammalinolenic acid
(DGLA) and arachidonic acid (AA) are the main EFA metabolites of
the n-6 series while eicosapentaenoic acid (EPA) and
docosahexaenoic acid (DHA) are the main metabolites of the n-3
series. DGLA, AA, EPA and DHA are important constituents of most of
the lipids in the body. As well as being important in themselves
they can also give rise to a wide range of oxygenated derivatives,
the eicosanoids, including the prostaglandins, leukotrienes and
other compounds. The fatty acids likely to be of particular value
in therapy are DGLA, AA, EPA and DHA, together with GLA, the
precursor of DGLA, stearidonic acid (SA), the precursor of EPA and
DPA (22:5n-3), the precursor of DHA, and adrenic acid.
Further there are fatty acids such as oleic acid, parinaric acid
and columbinic acid that are not EFAs but may have significant
effects in the body. One of the most interesting of these is
conjugated linoleic acid which as noted earlier has a range of
desirable effects.
It used to be thought that, both in nutrition and in therapy of
disease, it was sufficient to supply linoleic and alpha-linolenic
acids and the body's own metabolism would do the rest. It is now
widely accepted that this is not true. Different diseases may have
different abnormal patterns of EFAs and because of problems in
metabolism these cannot simply be corrected by giving linoleic or
alpha-linolenic acid. It is therefore appropriate in many
situations to provide increased amounts of one of the other EFAs or
to give two or more of the EFAs simultaneously. While the EFAs can
be supplied in various forms and in various mixtures, it is
convenient in both nutrition and in medical treatment to be able to
supply the fatty acids as particular molecules. Equally in various
situations it may be desirable to give the EFA or other fatty acid
in association with an amino acid, vitamin, drug or other molecule
which in itself has desirable properties.
To date, proposals for administration of two fatty acids
simultaneously have been in terms of particular triglycerides,
following the natural occurrence of essential fatty acids in
triglycerides form. However, triglycerides, unless symmetrical
about the 2-carbon, are chiral and that fact, coupled with acyl
migration between the alpha and beta positions makes the synthesis
of specific triglycerides a difficult task. Such migration may take
place after synthesis creating particular problems in a drug
regulatory context. The lack of specificity when two fatty acids
are present in the same triglyceride molecule creates many problems
in synthesis, pharmacology, formulation and stability. Moreover
triglycerides can be slow and difficult to synthesise. When treated
under similar conditions propane diol derivatives can be made much
more rapidly.
For purposes of convenient administration of different fatty acids
simultaneously or indeed of a single fatty acid in high amounts in
well tolerated form, use is thus desirably made of esters of
diols.
Chemical Nature of Bioactives which may be Derivatised According to
the Present Disclosure
The present specification covers fatty acid (or fatty alcohol)
derivatives of bioactives with an available carboxyl, alcohol or
amino group such that a single, well defined chemical entity is
formed. The coupling may be direct yielding bipartate compounds or
spaced with an appropriate link group, yielding tripartate
compounds, in terms of the number of moieties into which the
compounds split.
Classes of Bioactives by Chemistry
Among the classes of compounds are those below, where n is
conveniently 1 to 3. The substances claimed herein include diesters
of class (a) (ii); n=3. Also claimed are phosphate esters of class
(b) (iv); n=3. Substances where n is a greater or lesser number, or
where the links are not ester links, may be of value for similar
reasons and are disclosed but largely not claimed currently. (a)
Bioactives with a free carboxyl group--these may be derivatised as
follows: ##STR00002## In all of the above categories where n is
suitably 1 to 3, the carbon chain of the unsaturated fatty acid or
alcohol is represented by: ##STR00003## In all of these categories
"unsaturated fatty acid" (and the derived "unsaturated fatty
alcohol") represents a member of a group comprising oleic acid (and
oleoyl alcohol) and any fatty acid (or corresponding fatty alcohol)
with two or more cis or trans double bonds. However, the fatty
acids likely to be of most value in this context are the essential
fatty acids shown in FIG.1 and in particular GLA, DGLA, AA, SA, EPA
and DHA. For particular purposes conjugated linoleic acid and
columbinic acid may be of great interest.
General Discussion of Synthesis
The individual fatty acids may be purified from natural animal,
vegetable or microbial sources or may be chemically synthesized by
methods known to those skilled in the art or developed
hereafter.
The individual fatty alcohols may be prepared by chemical reduction
of the fatty acids outlined above by methods known to those skilled
in the art or developed hereafter.
Derivatisation of bioactives in classes (a), (b) and (c)
[subclasses (ii) and (iii)] requires the formation of one or more
ester bonds. Such chemistry may be achieved by any reasonable
method of ester synthesis and especially: (a) by reaction of
alcohol with acid chloride, acid anhydride or suitably activated
ester with or without the presence of an organic tertiary base,
e.g. pyridine, in a suitable inert solvent, e.g. dichloromethane,
at a temperature between 0.degree. and 120.degree. C. (b) by
reaction of alcohol with acid or acid, short or medium chain alkyl
ester, in the presence of a suitable acid catalyst, e.g. 4-toluene
sulfonic acid, with or without a suitable inert solvent, e.g.
toluene, at a temperature between 500 and 180.degree. C. such that
the water formed in the reaction is removed, e.g. under vacuum. (c)
by reaction of alcohol with acid in the presence of a condensing
agent, e.g. 1,3-dicyclohexylcarbodiimide, with or without the
presence of a suitable organic tertiary base, e.g.
4-(N,N-dimethylaminopyridine), in an inert solvent, e.g.
dichloromethane, at a temperature between 0.degree. and 50.degree.
C. (d) by reaction of alcohol with acid or acid, short or medium
chain alkyl ester, or acid, activated ester, e.g. vinyl, in the
presence of a hydrolase enzyme, e.g. hog liver esterase, with or
without a suitable solvent, e.g. hexane, at temperatures between
20.degree. and 80.degree. C. under conditions such that the water
or alcohol or aldehyde byproduct is removed, e.g. under vacuum. (e)
by reaction of acid with suitable alcohol derivative, e.g. iodide,
with or without the presence of a suitable base, e.g. potassium
carbonate, in a suitable inert solvent, e.g. dimethylformamide, at
a temperature between 0.degree. and 180.degree. C. (f) by reaction
of alcohol with acid, short or medium chain alkyl ester, in the
presence of a catalytic amount of an alkoxide of type M+OY-- where
M is an alkali or alkaline earth metal, e.g. sodium, and Y is an
alkyl group containing 14 carbon atoms which may be branched,
unbranched, saturated or unsaturated, with or without the presence
of a suitable solvent, e.g. toluene, at temperatures between
50.degree. and 180.degree. C. such that the lower alcohol, HOY, is
removed from the reaction mixture, e.g. under vacuum.
Derivatisation of bioactives in class (c) require the formation of
an amide bond. Such chemistry may be achieved by any reasonable
method of amide synthesis and especially: (g) by reaction of amine
with acid chloride, acid anhydride or suitably activated ester with
or without the presence of an organic tertiary base, e.g. pyridine,
in a suitable inert solvent, e.g dichloromethane, at a temperature
between 0.degree. and 120.degree. C. (h) by reaction of amine with
acid in the presence of a condensing agent, e.g.
1,3-dicyclohexylcarbodiimide, with or without the presence of a
suitable organic tertiary base, e.g. 4-(N,N-dimethylaminopyridine),
in an inert solvent, e.g. dichloromethane, at a temperature between
0.degree. and 50.degree. C. (i) by reaction of amine with acid or
acid, short or medium chain alkyl ester, or acid, activated ester,
e.g. vinyl, in the presence of a hydrolase enzyme, e.g. hog liver
esterase, with or without a suitable solvent, e.g. hexane, at
temperatures between 20.degree. and 80.degree. C. under conditions
such that the water or alcohol or aldehyde byproduct is removed,
e.g. under vacuum.
Derivatisation of bioactives in class (b) (iv) requires the
formation of phosphate ester bonds. Such chemistry may be achieved
by any reasonable method of phosphate ester synthesis and
especially: (j) by reaction of alcohol (e.g. UFA, 3-hydroxypropyl
ester) with a suitably activated phosphate derivative (e.g.
POCl.sub.3) with a tertiary base (e.g. Et.sub.3N) in a suitable
solvent (e.g. THF) at a temperature less than 10.degree. C. to
yield crude phosphorodichloridate. This is followed by reaction of
alcohol (e.g. .alpha.-tocopherol) with the crude
phosphorodichloridate with a tertiary base (e.g. Et.sub.3N) in a
suitable solvent (e.g. THF) at around ambient temperature to yield
crude phosphorochloridate. This may be hydrolysed (e.g. by addition
of water and Et.sub.3N) to yield phosphodiester. Alternatively,
addition of methanol yields a phosphotriester which may be
demethylated using a suitable nucleophile (e.g. lithium bromide) in
a suitable solvent (e.g. methyl ethyl ketone) to yield the
phosphodiester. (k) by reaction of phosphomonoester (e.g. phosphate
of UFA, 3-hydroxypropyl ester) with alcohol (e.g. choline) in the
presence of a condensing agent (e.g. 1,3-dicyclohexylcarbodiimide)
in a suitable solvent at a suitable temperature. (l)
transphosphatidylation reaction of
2-deoxy-2-lysophosphatidylcholine with primary or secondary alcohol
catalysed by phospholipase D.
In general the chemistry of course depends on the nature of the
compounds to be linked and on whether links are direct or indirect.
Fatty acid pairs may for example be linked directly as fatty
acid-fatty alcohol esters or as anhydrides, and if diol linkers are
used ether links to fatty alcohols are an alternative to the more
generally convenient ester links to fatty acids as such; in all
cases linking may again be by chemistry known in itself.
Examples of Pairs of Actives which may be Linked either Directly or
via a Link, Particularly a 1,3-Propane Diol Link
Examples of pairs of actives follow, the resulting compounds listed
being, to our knowledge, largely novel. So far as that is so, they
represent part of the invention as new chemical entities, as well
as being novel in use in treatment or prevention of disease,
whether or not currently claimed.
Fatty Acids
GLA-OA (OA=Oleic Acid), GLA-GLA, EPA-EPA, GLA-EPA, GLA-DHA, AA-DHA,
AA-EPA, GLA-AA, GLA-SA, SA-DHA, AA-SA, DGLA-DGLA, DGLA-GLA,
DGLA-SA, DGLA-AA, DGLA-EPA, DGLA-DHA, AA-AA, EPA-SA, EPA-DHA,
DHA-DHA, cLA-cLA, cLA-GLA, cLA-DGLA, cLA-AA, cLA-SA, cLA-EPA,
cLA-DHA, CA-CA, CA-GLA, CA-DGLA, CA-AA, CA-SA, CA-EPA, CA-DHA.
Vitamins
GLA-niacin, GLA-retinoic acid, GLA-retinol, GLA-pyridoxal,
Di-GLA-pyridoxine, di-EPA-pyridoxal and in general any of e.g. GLA,
DGLA, AA, SA, EPA or DHA with any vitamin including ascorbic acid,
Vitamin D and its derivatives and analogues, Vitamin E and its
derivatives and analogues, Vitamin K and its derivatives and
analogues, Vitamin B.sub.1 (thiamin), Vitamin B.sub.2 (riboflavin),
folic acid and related pterins, Vitamin B.sub.12, biotin and
pantothenic acid.
Amino Acids
GLA-tryptophan, GLA-proline, GLA-arginine, GLA- or
DHA-phenylalanine, GLA-GABA, GLA-aminolevulinic acid and in general
any of e.g. GLA, DGLA, AA, SA, EPA or DHA with any natural amino
acid or related compound such as taurine and carnitine.
Aromatic Acids
GLA-phenylbutyric acid, GLA-phenylacetic acid, GLA-trans-cinnamic
acid and in general any of e.g. GLA, DGLA, AA, SA, EPA or DHA with
any aryl alkanoic or aryl alkenoic acid.
Steroids
GLA-hydrocortisone, GLA-oestradiol, GLA- and
DHA-dehydroepiandrosterone and in general any of e.g. GLA, DGLA,
AA, SA, EPA or DHA with any natural or synthetic steroid, such as
any oestrogen, any progestin, any adrenal steroid and any
anti-inflammatory steroid, particularly betamethasone, prednisone,
prednisolone, triamcinolone, budesonide, clobetasol, beclomethasone
and other related steroids.
Anti-oxidants
GLA-lipoic acid, DHA-lipoic acid, GLA-tocopherol,
di-GLA-3,3'-thiodipropionic acid and in general any of e.g. GLA,
DGLA, AA, SA, EPA or DHA with any natural or synthetic anti-oxidant
with which they can be chemically linked. These include phenolic
anti-oxidants (e.g. eugenol, carnosic acid, caffeic acid, BHT,
gallic acid, tocopherols, tocotrienols and flavonoid anti-oxidants
(e.g. myricetin, fisetin)), polyenes (e.g. retinoic acid),
unsaturated sterols (e.g. .DELTA..sup.5-avenosterol), organosulfur
compunds (e.g. allicin), terpenes (e.g. geraniol, abietic acid) and
amino acid anti-oxidants (e.g. cysteine, carnosine).
Drugs
GLA and indomethacin, ibuprofen, fluoxetine, ampicillin, penicillin
V, sulindac, salicylic acid, metronidazole, fluphenazine, dapsone,
tranylcypromine, acetyl carnitine, haloperidol, mepacrine,
chloroquine, penicillin, tetracycyline, pravastatin,
bisphosphonates such as efidronic acid, pamidronic acid and
clordronic acid and their sodium salts, adenosylosuccinate and
adenylosuccinate and related compounds and agents used as x-ray
contrast media, and in general any of e.g. GLA, DGLA, AA, SA, EPA
or DHA with any drug, particularly any drug used in the treatment
of infections, inflammatory diseases, including various forms of
arthritis, cancer, cardiovascular, respiratory, dermatological,
psychiatric, neurological, muscular, renal, gastrointestinal,
reproductive and other diseases.
Concepts Applied to NSAIDs; Effectiveness Shown
As a particular example of the concepts discussed, we have prepared
derivatives of various non-steroidal anti-inflammatory drugs
(NSAIDS) and in particular the GLA-ester of indomethacin.
Indomethacin as a non-steroidal anti-inflammatory drug is believed
to have a primarily intracellular mechanism of action by inhibiting
the enzyme cyclo-oxygenase, which converts arachidonic acid to
pro-inflammatory prostaglandin metabolites.
Indomethacin is known to penetrate cells very poorly and so has to
be given in relatively large doses which can produce many side
effects, thus indomethacin-GLA was compared with indomethacin
itself for its ability to penetrate cells, using a normal
fibroblast line, a breast cancer line and a malignant melanoma
line.
The results are set out in EPA-0 675 103 and show that in all the
cell lines the intracellular level of indomethacin after incubation
with indomethacin is very low and is mainly detected only in trace
amounts. In contrast, again in all cell lines, incubation with
indomethacin-GLA resulted in very substantial amounts of both
indomethacin-GLA and free indomethacin being found within the
cells. These results show unequivocally that the GLA ester of
indomethacin penetrates cells effectively and is then deterified
intracellularly to provide free indomethacin, and that in view of
the many similarities between the cell membrane barrier and the
blood-brain and skin barriers, the indomethacin-GLA will also be
effective in accelerating the penetration of indomethacin through
these barriers. Such penetration, and breakdown to free the
actives, is to be expected with all the compounds set out
herein.
The Present Invention as Claimed
Aspects of the invention are set out in the claims herein, the main
claim referring to compounds, when for use in therapy wherein a 1,3
propane diol residue forms a link between residues R.sup.1 and
R.sup.2 where R.sup.1 is an acyl or fatty alcohol group derived
from a C.sub.12-30 preferably C.sub.16-30 fatty acid desirably with
two or more cis or trans double bonds, and R.sup.2 is hydrogen, or
an acyl or fatty alcohol group as R.sup.1 the same or different, or
any other nutrient, drug or other bioactive residue.
The compounds will generally be acid-function bearing actives
esterified directly to the diol residue but for example with a
fatty alcohol or other hydroxy-function bearing active, a
phosphate, succinate or other difunctional acid group may be
interposed between the R.sup.1 and/or R.sup.2 group and the
1,3-propane diol residue, particularly when R.sup.2 is a nutrient,
drug or other bioactive with a hydroxy or amino function.
The invention is also discussed broadly below, concerning a wide
range of actives releasable in the body.
While direct linkages of bioactives and fatty acids (classes
(a)[i], (b)[i] and (c)[i] are discussed above, the present
invention concerns primarily class (a)[ii], n=3 whereby bioactives,
which may themselves be fatty acids, are linked to fatty acids as
diesters of 1,3-propane diol and class (b)(iv), n=3 whereby
bioactives, which may themselves be fatty alcohols or
3-hydroxypropyl esters of fatty acids, are linked via a phosphate
linkage to a fatty acid monoester of 1,3-propane diol. This diol
may also be regarded as 2-deoxyglycerol and the corresponding
diesters as 2-deoxy-1,3-diglycerides. Compounds in class (b)(iv),
n=3 are also based on 1,3-propane diol and may be regarded as
2-deoxy-2-tysophospholipids. The compounds listed herein are almost
all new chemical entities or at least have never previously been
used in treatment of human or animal disease.
As a compound the diol used as a link is, broadly, disclosed in the
literature among many other diols but we have seen that its use in
therapy in the form of an essential fatty acid diester or as a
compound with an essential fatty acid at one position and a
bioactive (not being an essential fatty acid) at the other, is both
undisclosed and particularly significant. Indeed it offers a
favourable way to give a single fatty acid as the monoester or
diester if a completely defined compound is required, as there is
no chiral centre such as is present in glycerol 1(3)-monoesters and
in diglycerides (.alpha.,.beta. and 1,3 where the fatty acid at
position 1 is different from that at position 3), nor do positional
isomers exist. Further, apart from administering individual acids,
such mono and diesters may have value in pharmaceutical formulation
as emulsifiers. The 1,3-propane diol structure is close to the
glycerol of natural triglycerides and an effective and safe
delivery system. Moreover it allows ready and unequivocal synthesis
of defined compounds without the problems of acyl migration shown
in triglycerides and without complications by optical isomers. We
have for example shown that intravenous infusion and oral
administration of a 1,3 propane diol GLA/EPA diester emulsion leads
to rapid in vivo release of free GLA and EPA and to further
metabolism of the GLA to AA and of the EPA to DHA. Similarly,
GLA-GLA and EPA-EPA diesters, and niacin-GLA and indomethacin-GLA
diesters have been shown to be absorbed following oral
administration and to release their active moieties.
Furthermore, as far as we are aware, all of the 1,3-propane diol
derived compounds set out on pages 17, 18 and 19 are novel
compounds which have never before been described. The specific
diols of fatty acids listed, and the diols where a fatty acid drawn
from the list of GLA, DGLA, AA, SA, EPA, DHA, cLA and CA is present
at one position and at the other position is a vitamin, amino acid,
aromatic acid, steroid, anti-oxidant or other therapeutic drug, are
new substances.
The fatty acid diesters have a wide variety of possible uses. They
may be used as pharmaceuticals for the treatment or prevention of
diseases in which abnormalities of fatty acids have been
identified. They may be added to foods or added to or used as
nutritional supplements for those who require the particular fatty
acid for the treatment or prevention of diseases. They may also be
used in foods or pharmaceuticals for veterinary use. They may
further be used for skin care.
As advantages or in various particular aspects including those
currently in the claims herein, the invention provides: (i) A
convenient and safe way of administering, for therapeutic or
nutritional purposes, one or two unsaturated fatty acid moieties,
or one unsaturated fatty acid and one bioactive that is not a fatty
acid. (ii) A derivative, of a bioactive required to cross lipid
membranes in the body to exert its action whether in entry to a
cell or in passing the skin, blood-brain or other barrier, through
a 1,3-propane diol linkage to an essential fatty acid of the
natural n-6 or n-3 series and especially GLA or DGLA, AA, SA, EPA
or DHA or the related fatty acids cLA or CA. (iii) A fatty acid
derivative of a drug such that the drug and fatty acid are mutually
efficacious. (iv) A method of improving the transport of a drug
across lipid membranes in the body, characterised by the
administration of the drug in a form as above. (v) A method of
manufacture of a medicament for improved therapy involving
transport of a drug across lipid membranes in the body,
characterised by incorporating the drug in a medicament in a form
as above. (vi) A method of manufacture of a medicament for
delivering one or two fatty acids from the list in (ii) above or
for delivering one of those fatty acids in association with another
active agent.
Examples of specific compounds have been given earlier herein;
synthesis examples come later.
Efficacy and Uses Generally
Particular uses of particular groups of compounds are indicated
elsewhere herein but the usefulness, generally, of the 1,3-propane
diol diesters may be illustrated by the following: 1. Improved
tolerability of fatty acids. Apart from the triglycerides, most
forms in which fatty acids can be administered including free
acids, salts, ethyl esters and other glycerides cause some degree
of gastrointestinal intolerance as shown by nausea, vomiting and
diarrhoea. The propane diol diesters in animal studies in rats and
mice have been found to be extremely well tolerated. For example,
the GLA-GLA and GLA-EPA diesters have been given to rats and mice
at doses of up to 10 g/kg without any evidence or diarrhoea. This
shows that the diesters are a very acceptable way of delivering
biologically active fatty acids. 2. Reduced toxicity of drugs. The
non-steroidal anti-inflammatory drugs such as aspirin and
indomethacin are notorious for causing severe gastrointestinal
toxicity with ulceration of the stomach and intestines and bleeding
into the gastrointestinal tract. Doses of indomethacin known to
cause gastrointestinal ulceration (5-30 mg/kg) were given to fasted
rats either in the form of free indomnethacin or the same amount of
indomethacin in a 1,3-propane diol diester with GLA in the other
position. The animals were sacrificed after 24 hours and the whole
gastrointestinal tract examined for ulceration. Whereas extensive
ulceration was found in the animals treated with indomethacin
alone, little or no ulceration was found in the animals treated
with GLA-indomethacin. 3. Efficient delivery of a biologically
active form of a fatty acid. GLA was administered in the form of
either GLA-GLA or GLA-EPA and EPA was administered in the form of
GLA-EPA or EPA-EPA. The diesters were given either orally by gavage
or intravenously in the form of a 20% emulsion made using 2% of oat
galactolipid as an emulsifier in doses from about 0.1 to 2.0 g/kg.
Animals were killed after 1, 2, 4, 8 and 24 hours and plasma, red
cells and liver collected. The presence of the unmetabolised
diesters was identified by high pressure liquid chromatography. The
presence of fatty acids derived from the diesters and of
metabolites of those fatty acids was checked by lipid extraction of
the liver, plasma or red cells, by separation of that lipid
fraction into triglycerides, phospholipids, cholesterol esters and
free fatty acids by thin layer chromatography, by methylation of
the fatty acids derived from those separated fractions and by
analysis of those fatty acids using gas chromatography using
methods well described in standard texts. These experiments showed
that after oral administration around 10% of the diester
administered could be identified in the diester form. Most of the
GLA or EPA was found in the free fatty acid or phospholipid and to
a lesser extent in the cholesterol ester and triglyceride
fractions. Moreover, particularly in the phospholipid fractions the
metabolites of GLA, DGLA and arachidonic acid, and the metabolites
of EPA, docosapentaenoic acid and DHA, could be found in increased
amounts. These observations indicate that the fatty acids are
readily released from the diester form and are then further
metabolised into biologically active substances. Similar results
were obtained from intravenous administration of the diester except
that at one hour around 40% of the diester remained in the original
form and the free fatty acids were released, metabolised and
incorporated into other lipid fractions over the following 24
hours. It is possible that the unchanged diester forms may have
biological activity themselves. Linoleic acid in a 1,3-diglyceride
form has been found to have anticancer effects which were selective
against cancer but not normal cells and which were not shared by
other forms of linoleic acid (A. Matsuzaki et al, Cancer Res. 1989;
49: 5702-7). It is possible that this and perhaps other actions
require the delivery of two molecules of the fatty acid spaced as
in a 1,3-diglyceride. Similar spacing will be achieved by a
1,3-propane diol and there may therefore be a particular value in
the intravenous administration of some of the propane diol
derivatives which will ensure that the diol form circulates for
some time before its complete metabolism.
The fatty acids have a large number of desirable biological and
therapeutic activities which have been detailed in numerous
publications by the inventors and by others. Four of the fatty
acids, GLA, DGLA, SA and EPA share a rather broad spectrum of
effects which include: 1. Cardiovascular actions including
vasodilatation, lowering of blood pressure, inhibition of platelet
aggregation, lowering of triglyceride and LDL-cholesterol levels,
elevation of HDL-cholesterol levels and inhibition of smooth muscle
proliferation. 2. Anti-inflammatory actions including reduction of
formation of pro-inflammatory mediators such as cytokines, and of
eicosanoids derived from arachidonic acid, reduction of neutrophil
migration and the neutrophil respiratory burst, reduction of local
inflammatory responses, inhibition of inflammation in various
animal models such as uric acid induced inflammation and adjuvant
arthritis, and treatment of various inflammatory disorders such as
osteoarthritis and rheumatoid arthritis. 3. Immunomodulatory
functions including the damping down of excessive immune and
allergic responses in animal models such as experimental allergic
encephalomyelitis and uveitis, bronchial and cutaneous
hyper-reactivity in sensitised animals, leading to the concept that
they are of value in human diseases where excessive immune
responses play a role. 4. Respiratory actions including
bronchodilatation and inhibition of brochoconstrictor actions. 5.
Improvements in calcium balance with increased calcium absorption,
reduced calcium excretion, increased deposition of calcium in bones
and reduced ectopic deposition of calcium in tissues such as
arteries and kidneys. 6. Anticancer effects of three sorts,
selective cytotoxic damage and induction of apoptosis in cancer
cells but not in normal cells, inhibition of growth by reduction of
action of growth factors and interference with second messenger
systems required for growth, inhibition of metastasis by various
actions including increased expression of E-cadherins and
inhibition of proteolytic enzymes such as urokinases, lipoxygenase
and matrix metalloproteinases, and inhibition of cancer-associated
cachexia. 7. Actions on nerve cells including maintenance of normal
nerve membrane structure and function and the normal pre- and
post-synaptic actions of neurotransmitters.
These desirable actions mean that this group of fatty acids can be
used in the treatment of may different disorders including
cardiovascular disorders of many types, inflammatory disorders
including rheumatoid arthritis, osteoarthritis, ulcerative colitis
and Crohn's disease, respiratory disorders including asthma,
psychiatric disorders including schizophrenia, alcoholism,
attention deficit disorder, depression and Alzheimer's disease,
neurological disorders including multiple sclerosis and
Huntington's chorea, renal and urinary tract disorders including
various types of renal inflammatory disease and urinary calcium
stones, metabolic disorders including osteoporosis and ectopic
calcification, and gastrointestinal ulcerative and inflammatory
diseases. Although conjugated linoleic acid (cLA) has not been
nearly as widely tested as, say GLA or EPA, it also seems to have a
wide range of actions including effects valuable in the treatment
of cancer, cardiovascular and metabolic diseases.
GLA, DGLA, AA and columbinic acid have desirable actions on the
skin and are particularly valuable in the treatment of skin
diseases such as atopic eczema, psoriasis, urticaria and allergic
reactions.
AA is often regarded as a potentially harmful fatty acid. However,
it is an essential constituent of all normal cell membranes and has
been found to be present in low levels in various illnesses
including atopic eczema, schizophrenia (Horrobin et al,
Schizophrenia Res. 1994; 13: 194-207) and cardiovascular disorders
(Horrobin, Prostaglandins Leukotr. EFAs 1995; 53: 385-96). AA is
likely to be of particular value in these situations and also in
other psychiatric disorders such as alcoholism and attention
deficit disorder where levels are also often low.
DHA shares some of the above actions of the EFAs but is found in
particularly important amounts in cell membranes and especially in
the membranes of the heart, the retina and the brain. DHA also has
potent anti-inflammatory and desirable cardiovascular effects. DHA
is likely to be of particular value in cardiovascular disorders, in
retinal and visual disorders including retinitis pigmentosa, senile
macular degeneration and dyslexia, and in psychiatric and
neurological disorders including schizophrenia, attention deficit
disorder, depression, alcoholism, Alzheimer's disease and other
forms of dementia and multiple sclerosis.
Infections have also recently been identified as likely to respond
to fatty acids, especially to GLA and DGLA, EPA and DHA. Many
bacteria are killed by these fatty acids, including strains which
are highly resistant to antibiotics. Recent work from a number of
laboratories has also shown that these highly unsaturated fatty
acids are important in successful responses to diseases like
malaria and to protozoal diseases.
It is thus apparent that various specific fatty acids are likely to
be able to add to the efficacy of drugs and other bioactive
substances of almost any class, in both the treatment and
prevention of disease, in skin care and in nutrition, as well as
having valuable therapeutic effects when given in the diol form as
a single fatty acid or as two different fatty acids in the same
molecule. Of particular value in therapy is that under most
circumstances the fatty acids are remarkably non-toxic and can be
administered safely in large doses without the risk of important
side effects.
As a specific example of the therapeutic efficacy of the diesters,
the 1,3 GLA-EPA propane diol diester was tested in the treatment of
the ASPC-1 human pancreatic -cancer transplanted subcutaneously
into nude mice which because they lack thymus function are able to
accept foreign transplants without rejection. 15 mice were each
injected subcutaneously with 15 million ASPC-1 cells suspended in
Matrigel and DMEM buffer. In all animals a tumour developed whose
size could be measured using callipers and whose volume could be
estimated from the linear dimensions. Tumour size in each animal
was measured twice weekly for five weeks. The animals were divided
into three groups. 5 animals were used as controls and received 10
g/kg corn oil per day only. 5 animals received 10 g/kg corn oil per
day but in addition received two injections per week of a dose of
1.5 g/kg of the GLA-EPA diester. The diester was administered in
the form of a 20% emulsion in which 2% of oat galactolipid was used
as the emulsifier; the intravenous emulsion was very well tolerated
and caused no haemolysis or thrombophlebitis or any other form of
distress to the animals. The other 5 animals instead of the corn
oil received 10 g/kg/day of the GLA-EPA diester. The treatments
were continued for three weeks and then the tumours were allowed to
grow for a further two weeks before the animals were sacrificed and
the tumours excised and weighed. The mean tumour weights were:
control group, 1240.+-.290 mg; intravenous GLA-EPA group,
820.+-.180 mg; oral GLA-EPA group, 490.+-.160 mg. Tumour growth was
thus substantially inhibited by both oral and intravenous
administration of the GLA-EPA diester without causing any side
effects or distress in the animals. This demonstrates that the
GLA-EPA diester can be effectively used in the treatment of cancer
as would be predicted by the effects of GLA and EPA given
separately in being able selectively to kill human cancer cells in
culture in the laboratory. Thus the diesters are biologically
active ways of administering the various fatty acids. The diesters
can therefore be reasonably expected to exert the many desirable
effects of the fatty acids which have been noted in many
publications in the literature (e.g. Horrobin D F, ed., Omega-6
Essential Fatty Acids: Pathophysiology and Roles in Clinical
Medicine: Wiley-Liss, New York, 1990. Simopoulos A P et al, eds,
Health Effects of Omega-3 Polyunsaturated Fatty Acids in Seafoods,
Karger, Basel, 1991. Fats and Oils in Human Nutrition, World Health
Organization, Rome, 1994. Unsaturated Fatty Acids: Nutritional and
Physiological Significance. British Nutrition Foundation, Chapman
and Hall, London, 1992).
Specific Uses of Particular 1,3-Propane Diol Compounds
1. 1,3-propane diol as derivatives containing: two fatty acids in
which one fatty acid is GLA or DGLA and the other is GLA, DGLA, SA,
EPA, DHA, cLA (conjugated linoleic acid) or CA (columbinic acid)
for the treatment of: (a) complications of diabetes, particularly
neuropathy and retinopathy; and improvement of responses to insulin
in diabetes and pre-diabetes; (b) cancers; (c) osteoarthritis; (d)
rheumatoid arthritis; (e) other inflammatory and auto-immune
diseases including Sjogren's syndrome, systemic lupus, ulcerative
colitis, Crohn's disease and uveitis; (f) respiratory diseases
including asthma; (g) neurological disorders including multiple
sclerosis, Parkinson's disease and Huntington's chorea; (h) renal
and urinary tract disorders; (i) cardiovascular disorders; (j)
degenerative diseases of the eye including retinitis pigmentosa and
senile macular degeneration; (k) psychiatric disorders including
schizophrenia, Alzheimer's disease, attention deficit disorder,
alcoholism and depression; (l) prostatis hypertrophy and
prostatitis; (m) impotence and male infertility; (n) mastalgia; (o)
male pattern baldness; (p) osteoporosis; (q) dermatological
disorders, including atopic eczema, hand eczema, psoriasis,
urticaria and allergic disorders; (r) dyslexia and other learning
disabilities; (s) cancer cachexia.
2. 1,3-propane diol as derivatives containing two fatty acids in
which one fatty acid is AA and the other is AA, GLA, DHA, DGLA or
EPA for treatment of the disorders as at (1) above and especially
(a), (g), (i), (k), (q) and (r).
3. 1,3-propane diol as derivatives containing two fatty acids in
which one fatty acid is EPA and the other is EPA or DHA for the
treatment of any of the disorders as at (1) above but especially
(b), (c), (d), (e), (f), (g), (h), (i), (j), (k), (p), (r) and
(s).
4. 1,3-propane diol as derivatives in which one position is
occupied by a fatty acid drawn from GLA, DGLA, AA, SA, cLA, EPA or
DHA and the other position is occupied by an agent, selected from
the following list, whose chemical structure is such that it can be
linked to the 1,3-propane diol by one of the linkages described
herein: (a) tryptophan for the treatment of any disease but
particularly for psychiatric, neurological, behavioural, pain and
other disorders and especially depression, sleep and migraine; (b)
phenylalanine for the treatment of any disease, but especially
depression, multiple sclerosis and chronic fatigue syndrome; (c)
arginine for the treatment of any disease but particularly diseases
in which the production of nitric oxide is defective; (d) carnitine
or carnitine derivatives for the treatment of any disease but
especially muscle weakness, cardiac failure, chronic fatigue
syndrome, Alzheimer's disease, and peripheral neuropathies; (e) any
other amino acid or related substance for the treatment of any
disease or aminolevulinic acid or derivative thereof for the
treatment of any disease but especially cancers; (f)
adenylosuccinate or related substances for the treatment of any
disease but especially muscular dystrophy, cardiac failure, chronic
fatigue and Alzheimer's disease and other dementias; (g) aspirin,
salicylic acid, indomethacin, ibuprofen, or any other non-steroidal
anti-inflammatory drug for the treatment of any disease but
especially of inflammatory disorders of pain, of Alzheimer's
disease and other dementias and of any disease in which platelet
aggregation should be inhibited; (h) any antibiotic for the
treatment of any appropriate infectious disease but especially
tetracycline, clindamycin, minocycline, chlortetracycline and
erythromycin for the treatment of acne; (i) any anti malarial or
anti-protozoal drug for the treatment of any disease, but
especially chloroquine, mnepacrine, quinacrine and mefloquine for
the treatment of malaria, protozoal disorders, inflammatory
disorders and schizophrenia; (j) any antifungal drug for the
treatment of any disease but especially metronidazole and
antifungal imidazoles and nitroimidazoles and amphotericin for the
treatment of fungal infections of various types; (k) any
anti-inflammatory steroid for the treatment of any disease but
especially hydrocortisone and betamethasone for the treatment of
skin disorders and beclomethasone and budesonide for the treatment
of asthma. (l) any gonadal steroid for the treatment of any disease
but especially oestrogens and progestogens for the treatment of
ovarian deficiency and osteoporosis and androgens for the treatment
of testicular deficiency; (m) any adrenal steroid for the treatment
of any disease, but especially dehydroepiandrosterone for the
treatment of disorders associated with ageing; (n) any retinoid for
the treatment of any disease but especially tretinoin and
isotretinoin for the treatment of dermatological disorders and for
use in skin care; (o) any anticancer agent for the treatment of
cancer; (p) any antipsychotic agent for the treatment of
schizophrenia and other psychoses; (q) any antidepressive agent for
the treatment of any disease but especially for the treatment of
depression; (r) any anti-anxiety agent for the treatment of any
disease, but especially for the treatment of anxiety and panic
attacks; (s) any immunosuppressive agent for the treatment of any
disease but especially cyclosporine and tacrolimus for the control
of immunity after organ transplantation and for the treatment of
autoimmune and inflammatory disorders including psoriasis, eczema,
asthma, rheumatoid arthritis and inflammatory bowel disease; (t)
any proton pump inhibitor or H2 antagonist for the treatment of any
disease but especially diseases associated with excess gastric acid
production or reduced defences against gastric acidity; (u) any
diuretic for any disease, but especially for diseases associated
with fluid retention and hypertension; (v) any calcium antagonist
used for any disease but especially for cardiovascular diseases;
(w) any angiotensin converting enzyme inhibitor or angiotensin
antagonist used for any disease but especially for cardiovascular
diseases; (x) any beta-blocker used for any disease but especially
for cardiovascular disorders; (y) any antiepileptic drug used for
any disease, but especially phenytoin, carbamazepine, valproate,
ethosuximide, vigabatrin or lamotrigine for the treatment of
epilepsy; (z) any hypolipidaemic agent for the treatment of any
disease but especially fibrates and statins used for cholesterol
lowering and cholesterol modification; (aa) any oral hypoglycaemic
or insulin-sensitising agents used in the management of diabetes;
(bb) any bisphosphonates used in the management of osteoporosis,
Paget's disease or cancer; (cc) any contrast agents used in
radiology including diatrizoate compounds, iodipamide,
ioglycamates, iopanoates, iophendylate, iothalamate, ioxaglate,
metrizamide and related compounds; (dd) any peptide or protein for
use in the treatment of diseases for which the peptide or protein
itself is used, including insulin, calcitonin, erythropoietin and
other peptides; (ee) any vitamin used in the treatment of any
disease, or used in foods, nutritional supplements or food
additives as a way of providing the vitamin effectively; (ff) any
antioxidant used in the management of any disease, but especially
for those diseases in which antioxidants may be especially
beneficial including cardiovascular diseases, cancer and
inflammatory disorders and any antioxidant used as a food or other
preservative or as a component of a food, food additive or
nutritional supplement, (gg) any porphyrin chlorin or
bacteriochlorin-based drug especially tetralis (hydroxy phenyl)
derivatives thereof used in photodynamic therapy of cancers.
Ease of Synthesis
Synthesis of Trielycerides
The following considers the advantages of use of 1,3-propane diol
compared in particular to triglycerides.
Specifically, it is proposed that 1,3-propane diol be used in place
of glycerol in the esterification of fatty acids, especially where
only one type of fatty acid (e.g. gamma-linolenic acid) is to be
attached to the three-carbon chain "backbone". Although diesters
and triglycerides are chemically very similar, the manufacture of
di-esters can be carried out under very mild conditions, and in a
matter of hours. To manufacture triglycerides, either harsh
conditions are required, or fatty acid chlorides must be used, or
bio-catalysts (which require reaction times of several days) are
necessary.
A summary of triglyceride synthesis methods is: chemical reaction
with metals, metal-chlorides, or organic acids as catalyst; use of
fatty-acid chlorides; use of immobilised enzymes.
All processes using acids, metals, or metal chlorides as catalysts
are very similar and share a common list of advantages and
disadvantages. Many of the problems are inherent to the methods,
i.e; acidic conditions and high temperatures (140.degree. C. to
180.degree. C.). The p-TSA method probably exhibits the least
problems, as this is carried out under the mildest conditions
(140.degree. C.). Reaction of glycerol with fatty acid chlorides is
done under "cold" conditions, but toxic gases are evolved and the
reaction can go out of control if not monitored carefully. This
method also suffers from the fact that the fatty acid chlorides
themselves must first be manufactured; this additional step reduces
the overall efficiency of the process. A particular family of
enzymes, the lipases, can be used to catalyse the esterification
reaction under very mild conditions (e.g. at 60.degree. C.), and
are probably the catalysts of choice when polyunsaturated fatty
acids are being used. However, most enzymes interact most
effectively with the 1- and 3-positions of glycerol. Addition of
fatty acid to the 2-position is slow, and often dependant upon
"acyl migration", i.e. a fatty acid must first be attached to the
1- or 3-position, and then migrate to the 2-position, where it
remains attached. Thus, triglyceride synthesis reactions which are
catalysed by enzymes can take days to approach completion.
In theory, the same methods can be applied to the esterification of
1,3-propanediol as can be applied to glycerol. However, when it is
considered that enzymes catalyse preferentially the addition of
fatty acids to the 1- and 3-positions of glycerol, it is clear they
should be particularly effective when used to make diesters. This
is indeed the case, with reactions being completed in a matter of
hours and at temperatures which are even lower (e.g. 45.degree. C.
to 60.degree. C.) than those required for triglyceride synthesis.
After four hours free fatty acid can be absent, and after eight
hours the yield of diester can be in excess of 95%, the balance
being monoester.
A further complexity with specific triglyceride syntheses is the
presence within glycerol of both primary and secondary hydroxyl
groups and a prochiral centre at the central carbon atoms. These
problems may be solved by the use of carefully selected protecting
groups and by chiral synthesis. However, this results in multistep
syntheses with decreasing yield and increasing impurity levels at
each step. In contrast, however, 1,3-propane diol possesses only
primary hydroxyl groups and no prochiral centres. The synthesis is
consequently reduced to two steps maximum with improved overall
yield and decreased impurity levels.
In summary, the reaction which prepares diesters from
polyunsaturated fatty acids and 1,3-propane diol is faster, and can
be carried out under much milder conditions, than can the
corresponding triglyceride synthesis. This leads to a more
economical and less wasteful production process and minimises the
risk of reactants or products becoming altered or degraded during
processing.
Formulations
The compounds may be formulated in any way appropriate and which is
known to those skilled in the art of preparing pharmaceuticals,
skin care products or foods. They may be administered orally,
enterally, topically, parenterally (subcutaneously,
intramuscularly, intravenously), rectally, vaginally or by any
other appropriate route.
Like triglycerides, the 1,3-propane diol diesters, especially those
containing two fatty acids, may be readily emulsified using
phospholipid or particularly galactolipid emulsifiers. Such
emulsions are particularly useful for administration via oral,
enteral and intravenous routes.
For example, fatty acid (UFA) diesters occur as free flowing oils
and therefore can be formulated as follows:
1. Preparation of 20% Emulsion of Diester of GLA and EPA with
1,3-Propane Diol
Oral emulsions were prepared by high-pressure homogenisation. The
particle size distributions and the zeta potential of the resulting
emulsions were determined by dynamic light scattering at room
temperature. The particle size measurements were carried out at
room temperature (Zetasizer 4 Malvern Instruments Limited).
An oil-in-water emulsion (batch size 200 g) was prepared containing
the following ingredients:
TABLE-US-00002 Ingredients % Emulsifier (Galactolipid)* 2.00
Diester (GLA-EPA) 20.00 Ascorbyl Palmitate (AP) 0.02 Vitamin E 0.5
Water 100.00
The emulsifier-galactolipid was dispersed in the diester and
Vitamin E, AP and water were mixed. The oil phase was added to the
aqueous phase under a high shear mix (Ultraturrax) at speed 4, for
a few minutes. This pre-emulsion was then homogenised at 80 MPA and
at 50.degree. C. for 6 cycles (mini-Lab 8.30 H; APV Rannie AS,
Denmark). The emulsion formed has an average droplet size of 230
nm.
Anti-microbial preservatives--potassium sorbate, and flavour, can
also be added to the above oral emulsion.
2. Preparation of Intravenous 20% Emulsion of Diester of GLA and
EPA with 1,3-Propane Diol
In a similar manner, 200 g of an oil-in-water emulsion was prepared
containing the following ingredients:
TABLE-US-00003 Ingredients % Emulsifier 2.0 Diester (GLA-EPA) 20.0
Glycerol 2.0 Water add to 100.00
The above emulsion, homogenised for 6 minutes in a high pressure
homogeniser had an average droplet size of 211 nm, a zeta potential
of -40 mV. These I.V. emulsions can be either filtered through a
membrane with a pore size of 0.22 microns or can be autoclaved with
change in droplet size.
The doses of the actives to be administered largely range from 1 mg
to 200 g per day, preferably 10 mg to 10 g and very preferably 10
mg to 3 g, according to their kind. In the treatment of cancer
preferable doses may be in the 2-150 g/day range. They may be
administered topically where appropriate in preparations where the
actives form from 0.001% to 50% of the topical preparation,
preferably 0.05% to 20% and very preferably 0.1% to 10%.
EXAMPLES
Illustrative syntheses of NSAID's linked to fatty acids are given
in published EPA-0 675 103 referred to earlier. Illustrative
syntheses of the linking of fatty acids, through 1,3-propane diol
residues follow, with other generally illustrative material.
Example 1
1,3-(di-z,z,z-Octadeca-6,9,12-trienoyloxy)propane
(Diester of GLA with 1,3-Propane Diol)
A solution of 1,3-dicyclohexylcarbodiimide (1.07 g) and
4-(N,N-dimethylamino)pyridine (0.59 g) in methylene chloride (5 ml)
was added to a solution of 1,3-dihydroxypropane (0.152 ml) and
z,z,z-octadeca-6,9,12-trienoic acid (95%, 1.36 g) in methylene
chloride (15 ml). The reaction was stirred at room temperature
under nitrogen until it was complete as determined by tlc. Hexane
(80 ml) was added to the reaction. The precipitate was removed by
filtration and washed thoroughly with hexane. The combined
filtrates were concentrated and purified by flash chromatography to
yield 1,3-(di-z,z,z-octadeca-6,9,12-trienoyloxy)propane as a pale
yellow free flowing oil.
Example 2
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(z-octadeca-9-enoyloxy)propane
(Diester of GLA and Oleic Acid with 1,3-Propane Diol).
Part 1:
A solution of z,z,z-octadeca-6,9,12-trienoic acid (150 g) in
methylene chloride (500 ml) was added dropwise to a mixture of
1,3-dihydroxypropane (205 g), 1,3-dicyclohexylcarbodiimide (130 g)
and 4-(N,N-dimethylamino)pyridine (87 g) in methylene chloride
(2500 ml) at room temperature under nitrogen. When tlc indicated
that the reaction had gone to completion the reaction mixture was
filtered. The filtrate was washed with dilute hydrochloric acid,
water and saturated sodium chloride solution. The solution was
dried, concentrated and purified by dry column chromatography to
yield 1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane as a
pale yellow oil.
Part 2:
A solution of 1,3-dicyclohexylcarbodiimide (23.7 g) and
4-(N,N-dimethylamino)pyridine (15.9 g) in methylene chloride (200
ml) was added to a solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (33.6 g) and
z-octadeca-9-enoic acid (30 g) in methylene chloride (400 ml) under
nitrogen at room temperature. On completion of reaction as
evidenced by tic analysis the solution was diluted with hexane,
filtered, concentrated and purified by dry column chromatography to
yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(z-octadeca-9-enoyloxy)propane
as a free flowing pale yellow oil.
Example 3
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(z,z,z,z,z-eicosa-5,8,11,14,17-pen-
taenoyloxy)propane
(Diester of GLA and EPA with 1,3-Propane Diol).
Prepared as in Example 2, Part 2 but replacing z-octadeca-9-enoic
acid with z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoic acid.
Chromatography yielded
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(z,z,z,z,z-eicosa-5,8,11,-
14,17-pentaenoyloxy)propane as a pale yellow oil.
Example 4
1,3-di(z,z,z-Octadeca-6,9,12-trienoyloxy)propane
(Diester of GLA with 1,3-Propane Diol).
Prepared as in Example 2, Part 2 but replacing z-octadeca-9-enoic
acid with z,z,z-octadeca-6,9,12-trienoic acid. Chromatography
yielded 1,3-(di-z,z,z-octadeca-6,9,12-trienoyloxy)propane as a pale
yellow oil.
Example 5
(.+-.)-1-(1,2-Dithiolane-3-pentanoyloxy)-3-(z,z,z-octadeca-6,9,12-trienoyl-
oxy)propane
(Diester of Lipoic Acid and GLA with 1,3-Propane Diol)
A mixture of 1,3-dicyclohexylcarbodiimide (720 mg, 3.45 mmol) and
4-(N,N-dimethylamino)pyridine (480 mg, 3.98 mmol) in tert-butyl
methyl ester (15 ml) was added to a mixture of lipoic acid (645 mg,
3.12 mmol) and
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (1 g, 3
mmol) in tert-butyl methyl ether (30 ml). The mixture was stirred
at room temperature under nitrogen for 5 h, the progress of
reaction being monitored by tic (40% ethyl acetate/hexane). On
completion the mixture was filtered, concentrated and purified by
flash chromatography (hexane, 2% ethyl acetate/hexane, 5% ethyl
acetate/hexane and finally 10% ethyl acetate/hexane) to yield
(.+-.)-1-(1,2-dithiolane-3-pentanoyloxy)-3-(z,z,z-octadeca-6,9,12-trienoy-
loxy)propane as a viscous yellow oil.
Example 6
1-([Z]-5-Fluoro-2-methyl-1-[4-{methylsulfinyl}benzylidenedindene-3-acetylo-
xy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane
(Diester of Sulindac and GLA with 1,3-Propane Diol)
A solution of 1,3-dicyclohexylcarbodiimide (720 mg, 3.45 mmol) in
tert butyl methyl ether (30 ml) was added to a mixture of sulindac
(1.12 g, 3.15 mmol), 4-(N,N-dimethylamino)pyridine (480 mg, 3.9
mmol) and 1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (1
g, 3 mmol) in tert-butyl methyl ether (15 ml). The mixture was
stirred at room temperature under nitrogen for 5 h, the progress of
reaction being monitored by tlc (40% ethyl acetate/hexane). On
completion the mixture was filtered, concentrated and purified by
flash chromatography (40% ethyl acetate/hexane, then 50% ethyl
acetate/hexane and finally 60% ethyl acetate/hexane) to yield
1-([Z]-5-fluoro-2-methyl-1-[4-{methylsulfinyl}benzylidene]indene-3-acetyl-
oxy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane as a waxy yellow
solid.
Example 7
1-([R]-3-Acetoxy-4-[(trimethylammonio]butyroyloxy)-3-(z,z,z-octadeca-6,9,1-
2-trienoyloxy)propane
(Diester of Acetyl Carnitine and GLA with 1,3-Propane Diol).
Freshly distilled thionyl chloride (1.5 ml) was slowly added to
(R)-acetyl carnitine (1 g.) in a pear shaped flask. Care was taken
to contain the reagents at the bottom of the flask until a clear
solution resulted. After 4 hours at room temperature excess thionyl
chloride was removed under reduced pressure (keeping the flask
temperature less than 30.degree. C.). This yielded the acid
chloride as a highly hydroscopic white solid which was used
immediately without further purification. To the flask were added
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (1.4 g, 4.17
mmol) and dry THF (4 ml). The mixture was allowed to stand
overnight at room temperature. Tlc analysis (40% ethyl
acetate/hexane) indicated that the reaction had gone to completion.
The reaction mixture was added dropwise to hexane (250 ml) with
vigorous stirring. A fine off white precipitate formed which was
collected by centrifugation. On removal of the supernatant the
solid was resuspended in hexane and centrifuged. The hexane washing
procedure was carried out once more to yield
1-([R]-3-acetoxy-4-[trimethylammonio]butyroyloxy)-3-(z,z,z-octadeca-6,9,1-
2-trienoyloxy)propane.
Example 8
1-(3,3-Dimethyl-7-oxo-6-((phenoxyacetyl)amino]4-thia-1-azabicyclo[3.2.0]he-
ptan-2-oyloxy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane.
(Diester of Penicillin V and GLA with 1,3-Propane Diol).
A mixture of penicillin V (1 g, 2.9 mmol),
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (860 mg, 2.6
mmol), 1,3-dicyclohexylcarbodiimide (620 mg, 3 mmol) and
4-(N,N-dimethylamino)pyridine (catalytic amount) in dichloromethane
(30 ml) was stirred overnight at room temperature. The reaction
mixture was diluted with hexane (50 ml), filtered and concentrated
to dryness. The residue was washed with hexane (3.times.50 ml) to
remove unreacted
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane. The
semisolid residue was disolved in diethyl ether (150 ml), washed
with water (100 ml) and dried. The ether solution was diluted with
hexane (125 ml) and the solution filtered through a bed of silica
(4 cm.times.4 cm). The filtrate was concentrated, yielding
1-(3,3-dimethyl-7-oxo-6([phenoxyacetyl)amino]-4-thia-1-azabicyclo[3.2.0]h-
eptan-2-oyloxy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane as a
viscous colourless oil.
Example 9
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(I-(4-chlorobenzoyl)-5-methoxy-2-m-
ethyl-indole-3-acetyloxy)propane.
(Diester of Indomethacin and GLA with 1,3-Propane Diol).
A solution of 1,3-dicyclohexylcarbodiimide (58 g, 0.28 mol) and
4-(N,N-dimethylamino)pyridine (37.9 g, 0.31 mol) in methylene
chloride (800 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (79.5 g, 0.24
mol) and indomethacin (93.2 g, 0.26 mol) in methylene chloride (400
ml) at room temperature under nitrogen. Stirring was continued for
3 h. The mixture was filtered, concentrated and purified by dry
column chromatography (ethyl acetate/hexane). The product fractions
were pooled and concentrated, yielding
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-(1-(3-chlorobenzoyl)-5-methoxy-2-m-
ethyl-indole-3-acetyloxy)propane as a bright yellow viscous
oil.
Example 10
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(2-pyrrolidine
carboxy)propane
(Diester of Proline and GLA with 1,3-Propane Diol).
Part 1:
A solution of 1,3-dicyclohexylcarbodiimide (674 mg, 3.3 mmol) and
and 4-(N,N-dimethylamino)pyridine (472 mg, 3.9 mmol) in methylene
chloride (20 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (1 g, 2.97
mmol) and N-tBOC-proline (671 mg, 3.12 mmol) in methylene chloride
(20 ml) at room temperature under nitrogen. Stirring was continued
for 7 h and the mixture stored overnight at 0.degree. C. The
mixture was filtered and purified by column chromatography
(methanol/methylene chloride) to yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(N-tBOC-2-pyrrolidine
carboxy)propane as a yellow oil.
Part 2:
The protected product was dissolved in 10% v/v
anisole/trifluoroacetic acid (10 ml) and left at room temperature
under nitrogen for 30 minutes. After tlc analysis indicated that
deprotection was complete, the mixture was purified by column
chromatography (8% methanol/42% methylene chloride/50% ethyl
acetate) to yield
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-(2-pyrrolidine
carboxy)propane as a viscous orange oil.
Example 11
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(2-amino-3-indolylpropanoyloxy)pro-
pane
(Diester of Tryptophan and GLA with 1,3-Propane Diol).
Part 1:
A solution of 1,3-dicyclohexylcarbodiimide (674 mg, 3.3 mmol) and
and 4-(N,N-dimethylamino)pyridine (472 mg, 3.9 mmol) in methylene
chloride (20 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (1 g, 2.97
mmol) and N-TSOC-tryptophan (950 mg, 3.12 mmol) in methylene
chloride (20 ml) at room temperature under nitrogen. Stirring was
continued for 7 h and the mixture stored overnight at 0.degree. C.
The mixture was filtered and purified by column chromatography
(methanol/methylene chloride) to yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(N-tBOC-2-amino-3-indolylpropanoy-
loxy)propane as a yellow oil.
Part 2:
The protected product was dissolved in 10% v/v
anisole/trifluoroacetic acid (6.1 ml) and left at room temperature
under nitrogen for 15 minutes. After tlc analysis indicated that
deprotection was complete, the mixture was purified by column
chromatography (8% methanol/42% methylene chloride/50% ethyl
acetate) to yield
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-(2-amino-3-indolylpropanoyloxy)pro-
pane as a viscous red wax.
Example 12
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(.alpha.-amino-.beta.-phenyl-propi-
onyloxy)propane
(Diester of Phenylalanine and GLA with 1,3-Propane Diol).
Part 1:
A solution of 1,3-dicyclohexylcarbodiimide (1.77 g, 8.57 mmol) and
and 4-(N,N-dimethylamino)pyridine (1.24 g, 10.13 mmol) in methylene
chloride (30 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (2.62 g, 7.79
mmol) and N-TBOC-phenylalanine (2.17 g, 8.18 mmol) in methylene
chloride (30 ml) at room temperature under nitrogen. Stirring was
continued for 7 h and the mixture stored overnight at 0.degree. C.
The mixture was filtered and purified by column chromatography
(methanol/methylene chloride) to yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(N-tBOC-.alpha.-amino-.beta-
.-phenyl-propionyloxy)propane as a yellow oil.
Part 2:
The protected product was dissolved in 10% v/v
anisole/trifluoroacetic acid (17 ml) and left at room temperature
under nitrogen for 30 minutes. After tlc analysis indicated that
deprotection was complete, the mixture was purified by column
chromatography (8% methanol/42% methylene chloride/50% ethyl
acetate) to yield
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-(.alpha.-amino-.beta.-phenyl-propi-
onyloxy)propane as a viscous yellow oil.
Example 13
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(4-aminobutanoyloxy)propane
(Diester of GABA and GLA with 1,3-Propane Diol).
Part 1:
A solution of 1,3-dicyclohexylcarbodiimide (0.84 g, 4.06 mmol) and
and 4-(N,N-dimethylamino)pyridine (0.59 g, 4.79 mmol) in methylene
chloride (10 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (1.24 g, 3.96
mmol) and N-tBOC-GABA (0.75 g, 3.69 mmol) in methylene chloride (15
ml) at room temperature under nitrogen. Stirring was continued for
7 h and the mixture stored overnight at 0.degree. C. The mixture
was filtered and purified by column chromatography (ethyl
acetate/hexane) to yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(N-tBOC-4-amino
butanoyloxy)propane as a colourless oil.
Part 2:
The protected product was dissolved in 10% v/v
anisole/trifluoroacetic acid (10.5 ml) and left at room temperature
under nitrogen for 30 minutes. After tlc analysis indicated that
deprotection was complete, the mixture was purified by column
chromatography (8% methanol/42% methylene chloride/50% ethyl
acetate) to yield 1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-(4-amino
butanoyloxy)propane as a yellow oil.
Example 14
3,3'-thio-di-(1-Propionyloxy-(3-(z,z,z-octadeca-6,9,12-trienoyloxy)-propan-
e))
(Bis Diester of GLA and 1,3-Propane Diol with 3,3'-Thiodipropionic
Acid).
A solution of 1,3-dicyclohexylcarbodiimide (660 mg, 3.22 mmol) and
4-(N,N-dimethylamino)pyridine (445 mg, 3.64 mmol) in methylene
chloride (10 ml) was added with stirring to a solution of
1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane (940 mg, 2.8
mmol) and 3,3'-thiodipropionic acid (250 mg, 1.4 mmol) in methylene
chloride (30 ml) at room temperature under nitrogen. Stirring was
continued for 4 h. The mixture was diluted with hexane (50 ml),
filtered, concentrated and purified by flash chromatography (ethyl
acetate/hexane). The product fractions were pooled and concentrated
yielding
3,3'-thio-di-(1-propionyloxy-(3-(z,z,z-octadeca-6,9,12-trienoyloxy)-propa-
ne)) as a colourless oil.
Example 15
1-(1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-propyl)4-(z,z,z-octadeca-6,9,12-
-trienyl)butane-1,4-dioate
(Diester of (GLM Monoester with 1,3-Propane Diol) and GLA Alcohol
with Succinic Acid).
Part 1:
A mixture of 1-(z,z,z-octadeca-6,9,12-trienyloxy)-3-hydroxypropane
(10 g, 30 mmol) and succinic anhydride (3 g, 30 mmol) in dry THF
(100 ml) was stirred at room temperature until a clear solution
resulted. This solution was cooled to 0.degree. C. and a solution
of 1,8-diazabicyclo[5.4.0]undec-7-ene (4.5 ml, 30 mmol) in dry THF
(50 ml) added dropwise to it. After 3 h, tlc analysis indicated
that most of the monoester had reacted. A few more crystals of
succinic anhydride were added and stirring continued for a further
30 min. The reaction mixture was diluted with diethyl ether (250
ml) and washed with 2M hydrochloric acid (2.times.250 ml), water
(250 ml) and brine (250 ml). It was then dried (sodium sulfate) and
concentrated to dryness. The material was used without any further
purification.
Part 2:
Oxalyl chloride (3.9 ml, 45 mmol) was added to a solution of the
product from part 1 (13 g, 30 mmol) in methylene chloride (75 ml).
The mixture was stirred at room temperature under nitrogen for 2 h
and concentrated to dryness. Hexane (75 ml) was added and the
mixture concentrated to dryness. This process was repeated with two
further portions of hexane (75 ml ea.). The material was used
without any further purification.
Part 3:
A solution of the acid chloride prepared in part 2 (1 g 2.2 mmol)
in methylene chloride (10 ml) was added dropwise to a solution of
z,z,z-octadeca-6,9,12-trienol (635 mg, 2.4 mmol), triethylamine (1
ml, 7.2 mmol) and 4-(N,N-dimethylamino)pyridine (cat. amount) in
methylene chloride (20 ml) at room temperature. On completion of
reaction, the mixture was concentrated and purified by flash
chromatography (ethyl acetate/hexane) to yield
1-(1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-propyl)-4-(z,z,z-octadeca-6,9,-
12-trienyl)butane-1,4-dioate as a colourless oil.
Example 16
1-(2,3,5-Triiodobenzoyloxy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane
(Diester of 2,3,5-(Triiodobenzoic Acid and GLA with 1,3-Propane
Diol)
2,3,5-Triiodobenzoyl chloride (1.54 g, 3.08 mmol) was added to a
mixture of 1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane
(1 g, 2.97 mmol) and triethylamine (1 ml) in methylene chloride (80
ml) and the resulting mixture stirred overnight at room temperature
under nitrogen. The mixture was concentrated and purified by flash
chromatography (ethyl acetate/hexane) to yield
1-(2,3,5-triiodobenzoyloxy)-3-(z,z,z-octadeca-6,9,12-trienoyloxy)propane.
Example 17
(.+-.)-1-(1,2-Dithiolane-3-pentanoyloxy)-3-(z,z,z,z,z,z-docosa-4,7,10,3,16-
,19-hexaenoyloxy)propane
(Diester of DHA and Liptic Acid with 1,3-Propane Diol)
Part 1:
A solution of z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoic acid
(6.4 g, 19.5 mmol) in methylene chloride (225 ml) was added
dropwise to a solution of 1,3-propane diol (7.5 g, 99 mmol),
1,3-dicyclohexylcarbodiimide (4.65 g, 20 mmol) and
4-(N,N-dimethylamino)pyridine (2.1 g, 17 mmol) in methylene
chloride (225 ml) at -10.degree. C. The reaction mixture was
stirred overnight, warming up to room temperature. The reaction was
filtered, concentrated and purified by flash chromatography (ethyl
acetate/hexane) to yield
1-(z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoyloxy)-3-hydroxypropane
as a pale yellow oil.
Part 2:
A solution of 1,3-dicyclohexylcarbodiimide (720 mg, 3.45 mmol) and
4-(N,N-dimethylamino)pyridine (480 mg, 3.9 mmol) in methylene
chloride (30 ml) was added to a mixture of
1-(z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoyloxy)-3-hydroxypropane
(1.16 g, 3 mmol) and lipoic acid (645 mg, 3.12 mmol) and methylene
chloride (15 ml). After 2.5 h at room temperature under nitrogen
the mixture was filtered, concentrated and purified by flash
chromatography (ethyl acetate hexane) to yield
(.+-.)-1-(1,2-dithiolane-3-pentanoyloxy)-3-(z,z,z,z,z,z-docosa-4,7,10,13,-
16,19-hexaenoyloxy)propane as a yellow oil.
Example 18
Methyl-di(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-phosphate
Phosphotnester of 2 Molecules of 3-Hydroxypropyl Ester of GLA and 1
Molecule of Methanol)
Part 1:
Triethylamine (3.74 ml, 26.8 mmol) was added dropwise to a cooled
(0.degree. C.) solution of freshly distilled phosphorus oxychloride
(2.74 g, 17.9 mmol) in anhydrous THF (15 ml). To this mixture was
added dropwise a solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (5 g, 14.9
mmol) in anhydrous THF (15 ml). The temperature was kept at less
than 10.degree. C. throughout and the reaction kept under an
atmosphere of nitrogen. Tlc analysis after 15 min. indicated
complete disappearance of starting material. The mixture was
filtered and concentrated. Toluene (50 ml) was added and the
mixture concentrated. A further portion of toluene (50 ml) was
added and removed.
Part 2:
A solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (3 g, 9
mmol) in anhydrous THF (10 ml) was added dropwise to a solution of
crude phosphochloridate (7.5 mmol) (half of the batch prepared in
part 1 above) and triethylamine (3.2 ml, 22.5 mmol) in anhydrous
THF (20 ml) at room temperature under nitrogen. The reaction was
stored for 3 days at less than 10.degree. C. Methanol (15 ml) was
added and the reaction kept at room temperature until tlc indicated
complete reaction of the phoshorochloridate to form the desired
phosphotriester. Purification by flash chromatography (ethyl
acetate/hexane) yielded
methyl-di(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-phosphate as a
colourless oil.
Example 19
Di(z,z,z-Octadeca6,9,12-trienoyloxypropyl)phosphate.
(Phosphodiester of 2 Molecules of 3-Hydroxyproypl Ester of GLA)
Lithium bromide (104 mg, 1.13 mmol) in methyl ethyl ketone (1 ml)
was added to a solution of
methyl-di(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-phosphate (0.85
g, 1.13 mmol) (prepared as in example 18) in methyl ethyl ketone (1
ml) and the mixture was heated under reflux for 1 h. After cooling,
the mixture was dissolved in diethyl ether (3 ml) and extracted
with water (3 ml). Emulsions formed were broken by the addition of
a few drops of methanol. The organic layer was separated, dried
(sodium sulfate), concentrated and purified by flash chromatography
(methanol/chloroform) to yield
di(z,z,z-octadeca-6,9,12-trienoyloxypropyl)phosphate as a waxy
white solid.
Example 20
(2-Aminoethyl)-(z,z,z-octadeca-6,9,12-trienoyloxypropyl)phosphate
(Phosphodiester of Ethanolamine and 3-Hydroxypropyl Ester of
GLA)
Part 1:
A mixture of ethanolamine (0.5 ml, 8.25 mmol) and triethylamine
(4.2 ml, 30 mmol) in anhydrous THF (20 ml) was added to a solution
of crude phosphochloridate (7.5 mmol) (half of the batch prepared
in example 18, part 1 above) in anhydrous THF (20 ml), keeping the
temperature less than 10.degree. C. Progress of the reaction was
monitored by tlc. The mixture was stored for 3 days at less than
5.degree. C. After that time it was filtered, concentrated, diluted
with hexane (50 ml) and reconcentrated.
Part 2:
The product obtained from part 1 was dissolved in isopropanol (100
ml), acetic acid (10 ml) and water (40 ml) and the solution allowed
to stand under nitrogen at room temperature. When tic indicated
that the reaction had gone to completion the mixture was
concentrated and partitioned between acetonitrile (50 ml) and
hexane (50 ml). The hexane layer was separated, concentrated and
purified by flash chromatography (methanol/chloroform 1 water). The
pure fractions were pooled and concentrated. Addition of ethyl
acetate crashed out
(2-aminoethyl)-(z,z,z-octadeca-6,9,12-trienoyloxypropyl)phosphate
as a waxy cream coloured solid which was collected by
centrifugation.
Example 21
(z,z,z-Octadeca-6,9,12-trienoyloxypropyl)-(2-(N,N,N-trimethylammonium)ethy-
l)phosphate
(Phosphodiester of Choline and 3-Hydroxypropyl Ester of GLA).
Part 1:
A solution of 2-chloro-1,3,2-dioxaphospholane-2-oxide (430 mg, 3.4
mmol) in toluene (5 ml) was added to a cooled (0.degree. C.)
solution of 1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane
(1 g, 2.98 mmol) and tnethylamine (0.57 ml, 4.1 mmol) in toluene
(45 ml). The mixture was stirred overnight, warming up to room
temperature. Tlc analysis indicated that the reaction had not gone
to completion. Further portions of triethylamine (0.3 ml) and
2-chloro-1,3,2-dioxaphospholane-2-oxide (200 mg) (as a solution in
toluene (5 ml)) were added and the reaction allowed to continue for
a further overnight period. After that time tic indicated complete
reaction and the mixture was concentrated.
Part 2:
The crude product from part 1 was dissolved in acetonitrile (60
ml). A quarter of this solution (15 ml) and trimethylamine (10 ml)
were heated in a sealed tube at 60.degree. C. for 5 h (CAUTION).
The reaction was cooled and concentrated under a stream of nitrogen
to yield
(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-(2-(N,N,N-trimethylammonium)eth-
yl)phosphate.
Example 22
(z,z,z-Octadeca-6,9,12-trienoyloxypropyl)phosphate
(Phosphomonoester of 3-Hydroxypropyl Ester of GLA).
A solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (1.95 g, 5.8
mmol), pyridine (1.4 ml, 17.3 mmol) and anhydrous THF (15 ml) was
added dropwise with stirring to a cooled (0.degree. C.) solution of
phosphorus oxychloride (1.02 g, 6.6 mmol) in anhydrous THF (5 ml)
and the resultant mixture was kept at 0.degree. C. for 3 h. Aqueous
sodium bicarbonate (10% w/w, 10 ml) was added to the reaction
mixture. After stirring for 20 min. the mixture was poured into
ice/water (30 ml) and the solution acidified to pH 1 by the
dropwise addition of 2M hydrochloric acid. The mixture was
extracted with diethyl ether (2.times.30 ml). The ether extracts
were combined, dried and concentrated. The resultant oil was
azeotroped with dry pyridine to yield
(z,z,z-octadeca-6,9,12-trienoyloxypropyl)phosphate as a viscous
yellow oil.
Example 23
Methyl-(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-(.alpha.-tocopheryl)phosp-
hate
(Phosphotriester of .alpha.-Tocopherot, Methanol and
3-Hydroaypropyl Ester of GLA).
Part 1:
Triethylamine (7.5 ml) was added to a solution of freshly distilled
phosphorus oxychloride (1.26 g, 8.25 mmol) in anhydrous THF (7.5m)
at 0.degree. C. After 15 min. a solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (2.5 g, 7.5
mmol) in anhydrous THF (7.5 ml) was added dropwise over a period of
30 min. at 0.degree. C. Stirring at this temperature was continued
for 30 min. after the end of addition. .alpha.-Tocopherol (3.23 g,
7.5 mmol) in anhydrous THF (5 ml) was added dropwise at 10.degree.
C. and the resultant mixture was then stirred at 10.degree. C. for
1 h and then overnight, warming up to room temperature.
Part 2:
One quarter of the mixture prepared in part 1 above, triethylamine
(0.8 ml, 6 mmol) and methanol (10 ml) were stirred overnight under
nitrogen at room temperature. The reaction mixture was concentrated
and partitioned between ethyl acetate (30 ml) and water (20 ml)
with sodium chloride and methanol being added to break the
emulsion. The ethyl acetate layer was dried, concentrated and
purified by flash chromatography (chloroform) to yield
methyl-(z,z,z-octadeca6,9,12-trienoyloxypropyl)-(.alpha.-tocopheryl-
)phosphate.
Example 24
(z,z,z-Octadeca-6,9,12-trienoyloxypropyl)-(.alpha.-tocopheryl)phosphate
(Phosphodiester of .alpha.-Tocopherol and 3-Hydroxypropyl Ester of
GLA).
Triethylamine (2 ml) and water (5 ml) were added to one quarter of
the reaction mixture as prepared in example 23, part 1. The mixture
was stirred under nitrogen in an ice bath for 1 h, acidified to pH
1 with 2M hydrochloric acid and extracted into ethyl acetate (20
ml) and methanol (5 ml). The extract was dried concentrated and
purified by flash chromatography (chloroform) to yield
(z,z,z-octadeca-6,9,12-trienoyloxypropyl)-(.alpha.-tocopheryl)phosphate.
Example 25
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-5-(z,z,z,z,z-eicosa-5,8,11,14,17-pen-
taenoyloxy)pentane
(Diester of GLA and EPA with 1,5-Pentane Diol).
Part 1:
z,z,z-Octadeca-6,9,12-trienoyl chloride (2 g) was added dropwise to
a solution of 1,5-dihydroxypentane (3.5 g), triethylamine (0.94 ml)
and 4-(N,N-dimethylamino)pyridine (0.2 g) in methylene chloride (50
ml) with stirring at 0.degree. C. under nitrogen. On completion of
reaction as evidenced by tic the reaction mixture was washed with
dilute hydrochloric acid and water, dried and purified by column
chromatography yielding
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-5-hydroxypentane as a pale
yellow oil.
Part 2:
As for Example 2, Part 2 but replacing
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane with
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-5-hydroxypentane and
z-octadeca-9-enoic acid with
z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoic acid. Chromatography
yielded
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-5-(z,z,z,z,z-eicosa-5,8,11,14,17-pe-
ntaenoyloxy)pentane as a pale yellow oil.
Example 26
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-4-(z,z,z,z,z-eicosa-5,8,11,14,17-pen-
taenoyloxy)benzene
(Diester of GLA and EPA with 1,4-Dihydroxybenzene).
Prepared as in Example 25, Parts 1 and 2 but replacing
1,5-dihydroxypentane with 1,4-dihydroxybenzene in Part 1 and
replacing methylene chloride with tetrahydrofuran as the solvent in
Part 1. Chromatography yielded
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-4-(z,z,z,z,z-eicosa-5,8,11,14,17-pe-
ntaenoyloxy)benzene as a pale yellow oil.
Example 27
1,4-di(z,z,z-Octadeca-6,9,12-trienyl)-butane-1,4-dioate
(Diester of GLA Alcohol with Succinic Acid)
Part 1:
A solution of 1,8-diazabicyclo[5.4.0]undec-7-ene (0.54 ml) in dry
tetrahydrofuran (10 ml) was added dropwise to a cooled (0.degree.
C.) solution of z,z,z-octadeca-6,9,12-trienol (1 g) and succinic
anhydride (0.36 g) in dry tetrahydrofuran (20 ml). On completion of
reaction as evidenced by tlc, the reaction mixture was diluted with
diethyl ether and washed with dilute hydrochloric acid, water and
brine. The organic layer was dried, concentrated and used directly
in the second part of the reaction.
Part 2:
A solution of 1,3-dicyclohexylcarbodiimide (0.83 g) and
4-(N,N-dimethylamino)pyridine (0.55 g) in methylene chloride (20
ml) was added to a solution of
1-(z,z,z-octadeca-6,9,12-trienyl)-butane-1,4-dioate (1.32 g) and
z,z,z-octadeca-6,9,12-trienol (0.98 g) in methylene chloride (40
ml). On completion, as evidenced by tlc analysis, the reaction
mixture was diluted with hexane, filtered, concentrated and
purified by chromatography to yield
1,4-di(z,z,z-octadeca-6,9,12-trienyl)-butane-1,4-dioate as a pale
yellow oil.
Example 28
2-(2-Methyl-5-nitroimidazolyl)ether-z,z,z-octadeca-6,9,12-trienoate
(Ester of Metronidazole with GLA)
Method A:
To a suspension of metronidazole (206 g) in anhydrous acetonitrile
(2300 ml) and anhydrous pyridine (107 ml) was added with stirring
at room temperature under nitrogen z,z,z-octadeca-6,9,12-trienoyl
chloride (373 g) over a period of 30 mins. Shortly after the
addition of the acid chloride a clear solution was formed and
stirring was continued for 2 hours. The mixture was allowed to
stand overnight and the solvent was removed in vacua (50.degree.
C./20 mm Hg). To the residue was added ethyl acetate (1000 ml), any
precipitated solid being filtered off. The ethyl acetate solution
was washed successively with brine, 2M hydrochloric acid, saturated
aqueous sodium bicarbonate solution and finally brine. After drying
(sodium sulfate) the solvent was removed to give an orange oil.
This material was subjected to dry column chromatography giving
2-(2-methyl-5-nitroimidazolyl)ethyl-z,z,z-octadeca-6,9,12-trienoate
as a pale yellow, non-distillable oil.
Method B:
Metronidazole (1.9 g) was suspended in toluene (30 ml) and with
stirring the mixture was heated under reflux with a Dean and Stark
head for 20 mins. to remove any water present. To the boiling
solution was added, under nitrogen, z,z,z-octadeca-6,9,12-trienoyl
chloride (2.96 g) dropwise over a period of 20 mins. The mixture
was stirred and heated under reflux for a further 2 hours, giving a
dark reaction mixture. After cooling, this mixture was subjected to
dry column chromatography giving
2-(2-methyl-5-nitroimidazolyl)ethyl-z,z,z-octadeca-6,9,12-trienoate
as a pale yellow, non distillable oil.
Example 29
2-(2-Methyl-5-nitroimidazolyl)ethyl-z,z-octadeca-9,12-dienoate
(Ester of Metronidazole with LA)
To a suspension of metronidazole (1.9 g) in dry dichloromethane (20
ml) was added successively 4-(N,N-dimethylamino)pyridine (1.22 g),
1,3-dicyclohexylcarbodiimide (2.2 g) and linoleic acid (2.8 g). The
mixture was stirred at room temperature overnight. To the reaction
was added 2M hydrochloric acid (20 ml) and stirring was continued.
After filtration the organic layer was separated, washed with 50%
saturated brine and finally with saturated aqueous sodium
bicarbonate. The dichloromethane solution was dried (sodium
sulfate) and evaporated in vacuo (30.degree. C./20 mm Hg). To the
resulting residue was added petrol (bp 30-60.degree. C., 20 ml) and
the mixture allowed to stand at room temperature for 2 hours,
causing the precipitation of the remaining urea. This was removed
by filtration and the filtrate was applied to a dry column giving
2-(2-methyl-5-nitroimidazolyl)ethyl-z,z-octadeca-9,12-dienoate as a
pale yellow, non distillable oil.
Example 30
2-(2-Methyl-5-nitroimidazoloyl)ethyl-z,z,z-eicosa-8,11,14-trienoate
(Eter of Metronidazole with DGLA)
In a similar manner but replacing the linoleic acid with the
requisite amount of z,z,z-eicosa-8,11,14-trienoic acid there is
prepared
2-(2-methyl-5-nitroimidazoloyl)ethyl-z,z,z-eicosa-8,11,14-trienoate.
Example 31
2-(2-Methyl-5-nitroimidazoloyl)ethyl-z,z,z,z,z,z-docosa-4,7,10,13,16,19-he-
xaenoate
(Ester of Metronidazole with DHA)
In a similar manner but replacing the linoleic acid with the
requisite amount of z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoic
acid there is prepared
2-(2-methyl-5-nitroimidazoloyl)ethyl-z,z,z,z,z,z-docosa-4,7,10,1-
3,16,19-hexaenoate.
Example 32
4-[3-[2-(Trifluoromethyl)10H-phenothiazin-10-yl]]-1-piperazineethyl-z,z,z--
octadeca-6,9,12-trienoate
(Eter of Fluphenazine with GLA)
In a similar manner but replacing, the metronidazole with the
requisite amount of the free base of
4-[3-[2-(trifluoromethyl)-10H-phenothiazin-10-yl]]-1-piperazineethanol
(fluphenazine) and the linoleic acid with the requisite amount of
GLA there is prepared
4-[3-[2-(trifluoromethyl)-10H-phenothiazin-10-yl]]-1-piperazineethyl-z,z,-
z-octadeca-6,9,12-trienoate.
Example 33
4,4'-(bis z,z,z-Octadeca-6,9,12-trienoylamino) diphenylsulfone
(Bis Amide of Dapsone with GLA)
In a similar manner but replacing the metronidazole with the
requisite amount of 4,4'-diamino diphenylsulfone (dapsone) and the
linoleic acid with the requisite amount of GLA there is prepared
4,4'-(bis z,z,z-octadeca-6,9,12-trienoylamino)diphenylsulfone.
Example 34
N-Methyl-3-phenyl-3[.alpha.,.alpha.,.alpha.-trifluoro-p-tolyl]propyl;
z,z,z-Octadeca-6,9,12-trienamide
(Amide of Fluoxetine with GLA)
In a similar manner but replacing the metronidazole with the
requisite amount of
N-methyl-3-phenyl-3[.alpha.,.alpha.,.alpha.-trifluoro-p-tolyl]p-
ropylamine (fluoxetine) and the linoleic acid with the requisite
amount of GLA there is prepared
N-methyl-3-phenyl-3[.alpha.,.alpha.,.alpha.-trifluoro-p-tolyl]propyl-z,z,-
z-octadeca-6,9,12-trienamide.
Example 35
trans-1-(z,z,z-Octadeca-6,9,12-trienoylamino)-2-phenyl
cyclopropane
(Amide of Tranylcypromine with GLA)
In a similar manner but replacing the metronidazole with the
requisite amount of trans-1-amino-2-phenylcyclopropane
(tranylcypromine) and the linoleic acid with the requisite amount
of GLA there is prepared
trans-1-(z,z,z-octadeca-6,9,12-trienoylamino)-2-phenyl
cyclopropane.
Example 36
6-[(Aminophenylacetyl)amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]-
heptane-2-carboxylic Acid-z,z,z-octadeca-6,9,12-trienamide
(Amide of Ampicillin with GLA)
Triethylamine (0.3 ml) was added to a stirred suspension of
ampicillin (0.7 g) in anhydrous DMF (120 ml) under a nitrogen
atmosphere. To the resultant clear solution was added
z,z,z-octadeca-6,9,12-trienoic acid, N-hydroxysuccinimide ester
(0.75 g) while maintaining the reaction at 0-10.degree. C. The
reaction was stirred at this temperature for an additional hour
before allowing the mixture to stand at room temperature overnight.
Tlc analysis (40% THF/hexane) at this point indicated that most of
the succinimide ester had reacted. Water (40 ml) was added to the
reaction flask and the contents stirred. The solution was then
neutralised and extracted with ethyl acetate. The extract was
washed with water, dried (sodium sulfate) and concentrated to
dryness leaving the crude product as a yellow glass. Trituration
with hexane yielded 6-[(aminophenylacetyl)
amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic
acid-z,z,z-octadeca-6,9,12-trienamide as a yellow powder.
Example 37
z,z,z-Octadeca-6,9,12-trienyl-z,z,z-octadeca-6,9,12-trienoate
(Ester of GLA with GLA Alcohol)
1,3-dicyclohexylcarbodiimide (0.82 g) and
4-(N,N-dimethylamino)pyridine (0.48 g) in methylene chloride (5 ml)
were added to a solution of z,z,z-octadeca-6,9,12-trienol (0.95 g)
and z,z,z-octadeca-6,9,12-trienoic acid (1 g) in methylene chloride
(10 ml) with stirring at room temperature under nitrogen. On
completion of reaction as evidenced by tlc, hexane was added to the
reaction mixture which was subsequently filtered and purified by
column chromatography to yield
z,z,z-mtadeca-6,9,12-trienyl-z,z,z-octadeca-6,9,12-trienoate as a
pale yellow oil.
Example 38
z,z,z-Octadeca-6,9,12-trienyl-z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoate
(Ester of EPA with GLA Alcohol)
Prepared as in Example 37 but replacing
z,z,z-octadeca-6,9,12-trienoic acid with
z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoic acid.
Example 39
2-Methyl-3-(z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoyloxy)-4-formyl-5-(z,z,z-
,z,z-eicosa-5,8,11,14,17-pentaenoyloxy)methyl pyridine
(diEPA Ester of Pyridoxal)
To a suspension of pyridoxal hydrochloride (1.0 g) in methylene
chloride (20 ml) was added triethylamine (2.0 ml). A clear yellow
solution developed. With cooling in ice,
z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoyl chloride (1.73 g)
(prepared by reaction of EPA with oxalyl chloride in methylene
chloride). The mixture was stirred overnight under nitrogen,
warming up to room temperature. After dilution with an equal volume
of methylene chloride, the mixture was extracted with 2M
hydrochloric acid (20 ml), washed with water (3.times.20 ml), dried
and concentrated. Purification by flash chromatography (ethyl
acetate/hexane) yielded
2-methyl-3-z,z,z,z,z-eicosa-5,8,11,14,17-pentaenoyloxy-4-formyl-5-(z,z,z,-
z,z-eicosa-5,8,11,14,17-pentaenoyloxy) methyl pyridine as a clear
oil.
Example 40
2-Methyl-3-hydroxy-4-formyl-5-(z,z,z-octadeca-6,9,12-trienoyloxy)methyl
pyridine
(GLA Ester of Pyridoxal)
A solution of z,z,z-octadeca-6,9,12-trienoyl chloride (800 mg, 2.7
mmol) in methylene chloride (10 ml) was added slowly dropwise to a
mixture of pyridoxal hydrochloride (500 mg, 2.45 mmol),
triethylamine (1 ml, 7.2 mmol) and 4-(N,N-dimethylamino)pyridine
(few mg, catalytic amount) in methylene chloride (20 ml) at
0.degree. C. under nitrogen. On completion, as indicated by tlc,
the mixture was concentrated and purified by flash chromatography
(ethyl acetate/hexane), yielding
2-methyl-3-hydroxy-4-formyl-5-(z,z,z-octadeca-6,9,12-trienoyloxy)methyl
pyridine as a colourless oil which subsequently solidified.
Example 41
2-Methyl-3-hydroxy-4,5-di(z,z,z-octadeca-6,9,12-trienoyloxy)methyl
Pyridine
(Bis GLA Ester of Pyridoxine)
A solution of z,z,z-octadeca-6,9,12-trienoyl chloride (650 mg, 2.2
mmol) in methylene chloride (10 ml) was added slowly dropwise to a
mixture of pyridoxine hydrochloride (206 mg, 1 mmol), triethylamine
(0.7 ml, 5 mmol) and 4-(N,N-dimethylamino)pyridine (few mg,
catalytic amount) in methylene chloride (20 ml) at 0.degree. C.l
under nitrogen. On completion, as indicated by tic, (4 h) the
mixture was concentrated and purified by flash chromatography
(ethyl acetate/hexane), yielding
2-methyl-3-hydroxy-4,5-di(z,z,z-octadeca-6,9,12-trienoyloxy)methyl
pyridine as a colourless oil.
Example 42
1-(2-(2-Methyl-5-nitroimidazoloyl)ethyl)-4-(z,z,z-octadeca-6,9,12-trienyl)-
butane-1,4-dioate
(Diester of Metronidazole and GLA Alcohol with Succinic Acid)
A solution of 1,3-dicyclohexylcarbodiimide (780 mg, 3.8 mmol) and
4-(N,N-dimethylamino)pyridine (530 mg, 4.3 mmol) in methylene
chloride (15 ml) was added with stirring to a solution of GLA
alcohol succinate monoester (1.25 g, 3.3 mmol) (prepared as in
Example 27, part 1) and metronidazole (620 mg, 3.6 mmol) in
methylene chloride (30 ml) at room temperature under nitrogen. On
completion of reaction, as indicated by tlc, the mixture was
diluted with hexane, filtered, concentrated and purified by flash
chromatography (ethyl acetate/hexane). The product fractions were
pooled and concentrated, yielding
1-(2-(2-methyl-5-nitroimidazoloyl)ethyl)-4-(z,z,z-octadeca-6,9,12-trienyl-
)-1,4-butanedioate as a colourless oil.
Example 43
trans-1-(z,z,z-Octadeca-6,9,12-trienyloxycarbonylbutyloxyamino)-2-phenyl
Cyclopropane
(Succinic Acid, 1-GLA Alcohol Ester, 4-Tranylcypromine Amide)
A solution of 1,3-dicyclohexylcarbodiimide (315 mg, 1.52 mmol) and
4-(N,N-dimethylamino)pyridine (210 mg, 1.72 mmol) in methylene
chloride (10 ml) was added with stirring to a solution of GLA
alcohol succinate monoester (500 mg, 1.32 mmol) (prepared as in
Example 27, part 1) and tranylcypromine (225 mg, 1.32 mmol) in
methylene chloride (20 ml) at room temperature under nitrogen. On
completion of reaction, as indicated by tlc, the mixture was
diluted with hexane, filtered, concentrated and purified by flash
chromatography (ethyl acetate/hexane). The product fractions were
pooled and concentrated, yielding
trans-1-(z,z,z-octadeca-6,9,12-trienyloxycarbonyl
butyloxyamino)-2-phenyl cyclopropane as a colourless oil.
Example 44
(.+-.)-2,5,7,8-tetramethyl-2-(4',8',12'-trimethyldecyl)-6-chromanyl-z,z,z--
octadeca-6,9,12-trienoate
(GLA ester of .alpha.-Tocopherol).
z,z,z-Octadeca-6,9,12-trienoyl chloride (2.96 g, 10 mmol) was added
dropwise with stirring over the course of 2-3 minutes to a solution
of (.+-.)-.alpha.-tocopherol (4.3 g, 10 mmol) and pyridine (0.885
ml, 11 mmol) in methylene chloride (35 ml) under nitrogen at
-5.degree. C. The reaction was then stirred overnight, warming up
to room temperature. Tlc analysis showed that the reaction had gone
substantially towards completion. The reaction mixture was washed
with water (100 ml), 2M hydrochloric acid (10 ml in 100 ml water)
and water (4.times.100 ml). The organic layer was dried (sodium
sulfate) and concentrated. Purification by flash chromatography
(ether/hexane) yielded
(.+-.)-2,5,7,8-tetramethyl-2-(4',8',12'-trimethyldecyl)-6-chromanyl-z,z,z-
-octadeca-6,9,12-trienoate as a pale yellow oil.
Example 45
Androst-5-en-17-one-3-(z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoate)
(DH Ester of Dehydroepiandrosterone)
To a cooled (0.degree. C) mixture of dehydroepiandrosterone (1 g)
and triethylamine (1 ml) in methylene chloride (20 ml) was added
z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoyl chloride (1.33 g)
(prepared by reaction of DHA with oxalyl chloride in methylene
chloride). The mixture was stirred overnight, warming up to room
temperature. It was diluted with methylene chloride (20 ml),
extracted with 2M hydrochloric acid (20 ml), washed with water
(2.times.20 ml), dried and concentrated. Purification by flash
chromatography (ethyl acetate/hexane) yielded
dehydroepiandrost-5-en-17-one-3
(z,z,z,z,z,z-docosa-4,7,10,13,16,19-hexaenoate) as a clear oil.
Example 46
z,z,z-Octadeca-6,9,12-trienyl-(2-(z,z,z-octadeca-6,9,12-trienyloxy)acetate-
)
Diester of GLA and GLA Alcohol with Glycolic Acid.
Part 1:
A solution of chloroacetyl chloride (0.4 ml, 5 mmol) in methylene
chloride (10 ml) was added dropwise to a solution of
z,z,z-octadeca-6,9,12-trienol (1 g, 3.8 mmol) and triethylamine
(1.4 ml, 10 mmol) in methylene chloride (20 ml) at 0.degree. C.
Progress of reaction was monitored by tlc. After 3 h the reaction
had gone substantially but not completely towards completion. A few
more drops of chloroacetyl chloride were added. Tlc analysis within
5 minutes showed the reaction to be complete. The mixture was
washed with water (2.times.50 ml) and brine (50 ml), dried (sodium
sulfate) and concentrated. Toluene (50 ml) was added to remove
azeotropically last traces of water. This yielded the chloroacetyl
ester of GLA alcohol as a dark brown oil which was used without
further purification.
Part 2:
A mixture of z,z,z-octadeca-6,9,12-trienoic acid (700 mg, 2.5 mmol)
and cesium carbonate (410 mg, 1.25 mmol) was swirled in methanol
until a clear solution resulted. The mixture was then concentrated
and kept at 40.degree. C. under high vacuum for 1 h. This yielded
the cesium salt of GLA which was used without further
purification.
Part 3:
To the flask containing the cesium salt of GLA as prepared in part
2, was added the chloroacetyl ester of GLA alcohol (part 1) (500
mg, 1.5 mmol) and dry DMF (15 ml). The reaction was stirred under
nitrogen at room temperature. After 90 minutes, tlc analysis showed
the reaction to be complete. The reaction mixture was extracted
with hexane (2.times.40 ml) and the hexane extract washed with
brine (2.times.50 ml) and water (50 ml), dried (sodium sulfate) and
concentrated to yield
z,z,z-octadeca-6,9,12-trienyl-(2-(z,z,z-octadeca-6,9,12-trienyloxy)acetat-
e) as a colourless oil.
Example 47
Hydrocortisone-21-(z,z,z-octadeca-6,9,12-trienoate)
(GLA Ester of Hydrocortisone)
A solution of z,z,z-octadeca-6,9,12-trienoyl chloride (450 mg, 1.52
mmol) in methylene chloride (10 ml) was added slowly dropwise to a
mixture of hydrocortisone (500 mg, 1.38 mmol), triethylamine (420
.mu.l, 3 mmol) and 4-(N,N-dimethyl amino)pyridine (few mg,
catalytic amount) in methylene chloride (20 ml) at 0.degree. C.
under nitrogen. Tlc analysis after 4 h indicated that the reaction
had gone to completion. The mixture was concentrated and purified
by flash chromatography (ethyl acetate/hexane), yielding
hydrocortisone-21-(z,z,z-octadeca-6,9,12-trienoate) as a colourless
oil.
Example 48
z,z,z-Octadeca-6,9,12-trienyl-(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methylin-
dole-3-acetyloxy)acetate)
Diester of Indomethacin and GLA Alcohol with Glycolic Acid.
Part 1:
A mixture of indomethacin (895 mg, 2.5 mmol) and cesium carbonate
(410 mg, 1.25 mmol) was swirled in methanol until a clear solution
resulted. The solution was then concentrated and kept at 40.degree.
C. under high vacuum for 1 h. This yielded the cesium salt of
indomethacin as a bright yellow solid.
Part 2:
To the flask containing the cesium salt of indomethacin as prepared
in part 1 was added the chloroacetyl ester of GLA alcohol (prepared
as in Example 46 (part 1) (500 mg, 1.5 mmol) and dry DMF (15 ml).
The reaction was stirred under nitrogen at room temperature,
progress of reaction being monitored by tlc. After an overnight
period in the fridge, tlc analysis showed the reaction to be
complete. The mixture was partitioned between water (50 ml) and
ethyl acetate (50 ml). A few ml of brine were added to break the
emulsion. The ethyl acetate layer was washed with water (3=ml),
dried (sodium sulfate), filtered through a pad of silica and
concentrated to yield
z,z,z-octadeca-6,9,12-trienyl-(2-(1-(4-chlorobenzoyl)-5-methoxy-2-methyli-
ndole-3-acetyloxy)acetate) as a bright yellow oil.
Example 49
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(4-phenylbutanoyloxy)propane
Diester of 4-Phenylbutanoic Acid and GLA with 1,3-propane
Diol).
A solution of 1,3-dicyclohexylcarbodiimide (710 mg, 3.45 mmol) and
4-(N,N-dimethylamino)pyridine (475 mg, 3.9 mmol) in methylene
chloride (10 ml) was added to a solution of
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-hydroxypropane (1 g, 3
mmol) and 4-phenylbutanoic acid (520 mg, 3.15 mmol) in methylene
chloride (15 ml). The resultant mixture was stirred at room
temperature under nitrogen until it was complete as indicated by
tlc. The mixture was filtered, concentrated and purified by flash
chromatography to yield
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(4-phenylbutanoyloxy)propane.
Example 50
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(phenylacetoxy)propane
(Diester of Phenylacetic Acid and GLA with 1,3-Propane Diol)
In a similar manner to Example 49 but replacing the
4-phenylbutanoic acid with phenylacetic acid (430 mg, 3.15 mmol)
was prepared
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(phenylacetoxy)propane.
Example 51
1-(z,z,z-Octadeca-6,9,12-trienoyloxy)-3-(trans-cinnamoyloxy)propane
(Diester of trans-Cinnamic Acid and GLA with 1,3-Propane Diol)
In a similar manner to Example 49 but replacing the
4-phenylbutanoic acid with trans-cinnamic acid (470 mg, 3.15 mmol)
was prepared
1-(z,z,z-octadeca-6,9,12-trienoyloxy)-3-(trans-cinnamoyloxy)propane.
* * * * *