U.S. patent application number 11/989004 was filed with the patent office on 2010-02-04 for fatty acid analogues for equilibrating bone mineral density.
Invention is credited to Rolf Berge.
Application Number | 20100029973 11/989004 |
Document ID | / |
Family ID | 35295502 |
Filed Date | 2010-02-04 |
United States Patent
Application |
20100029973 |
Kind Code |
A1 |
Berge; Rolf |
February 4, 2010 |
Fatty acid analogues for equilibrating bone mineral density
Abstract
The invention comprises the use of compounds comprising non
.beta.-oxidizable fatty acid entities according to formula (I) or
(II) for the preparation of a pharmaceutical composition for the
prevention and/or treatment of conditions associated with
low/decreased bone mineral density (BMD), and/or for increasing the
BMD by decreasing the bone resorption.
Inventors: |
Berge; Rolf; (Bones,
NO) |
Correspondence
Address: |
Christopher Aniedobe;Dobe Law Group
7207 Hanover Pkwy ste. cd
Greenbelt
MD
20770
US
|
Family ID: |
35295502 |
Appl. No.: |
11/989004 |
Filed: |
July 10, 2006 |
PCT Filed: |
July 10, 2006 |
PCT NO: |
PCT/NO2006/000262 |
371 Date: |
May 6, 2009 |
Current U.S.
Class: |
558/177 ;
560/129; 560/147; 560/205; 562/26; 562/899; 564/123; 568/840 |
Current CPC
Class: |
A61P 19/08 20180101;
A61K 31/28 20130101; H05B 41/2885 20130101; A61K 31/19 20130101;
A61K 31/20 20130101; H05B 41/042 20130101; A61P 19/10 20180101;
Y02B 20/202 20130101; H05B 41/2882 20130101; A61K 31/00 20130101;
Y02B 20/00 20130101; H05B 41/2881 20130101 |
Class at
Publication: |
558/177 ;
560/205; 560/129; 560/147; 568/840; 564/123; 562/26; 562/899 |
International
Class: |
C07F 9/09 20060101
C07F009/09; C07C 69/52 20060101 C07C069/52; C07C 69/00 20060101
C07C069/00; C07C 31/00 20060101 C07C031/00; C07C 233/00 20060101
C07C233/00; C07C 327/00 20060101 C07C327/00; C07C 391/00 20060101
C07C391/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 18, 2005 |
NO |
20053519 |
Claims
1-10. (canceled)
11. A pharmaceutical composition for the prevention and/or
treatment of conditions associated with low/decreased bone mineral
density (BMD) comprising compound represented by the general
formula (I), ##STR00005## wherein R1, R2, and R3 represent i) a
hydrogen atom; or ii) a group having the formula CO--R wherein R is
a linear or branched alkyl group, saturated or unsaturated,
optionally substituted, and the main chain of said R contains from
1 to 25 carbon atoms; or iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer from 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group consisting of an oxygen atom, a sulphur atom, a
selenium atom, a CH.sub.2 group, a SO group, and a SO.sub.2 group;
iv) an entity selected from the group comprising
--PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a compound represented by the general formula
R''--COO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom,
a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer from 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; and R'' is a hydrogen atom or an alkyl group
containing from 1 to 4 carbon atoms; or a salt, prodrug or complex
of said compounds.
12. A pharmaceutical composition for the prevention and/or
treatment of conditions associated with low/decreased bone mineral
density (BMD) comprising compound represented by the general
formula (II), ##STR00006## wherein A1, A2 and A3 are chosen
independently and represent an oxygen atom, a sulphur atom or an
N-R4 group in which R4 is a hydrogen atom or a linear or branched
alkyl group, saturated or unsaturated, optionally substituted,
containing from 1 to 5 carbon atoms; wherein R1, R2, and R3
represent i) a hydrogen atom or a linear or branched alkyl group,
saturated or unsaturated, optionally substituted, containing from 1
to 23 carbon atoms; or ii) a group having the formula CO--R in
which R is a linear or branched alkyl group, saturated or
unsaturated, optionally substituted, and the main chain of said R
contains from 1 to 25 carbon atoms; or iii) a group having the
formula CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur
atom, a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group
or a SO.sub.2 group; n is an integer from 0 to 11; and R' is a
linear or branched alkyl group, saturated or unsaturated,
optionally substituted, wherein the main chain of said R' contains
from 13 to 23 carbon atoms and optionally one or more heterogroups
selected from the group consisting of an oxygen atom, a sulphur
atom, a selenium atom, a CH.sub.2 group, a SO group and a SO.sub.2
group; iv) an entity selected from the group comprising
--PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a salt, prodrug or complex of said compound.
13. The pharmaceutical composition of claim 11 or 12, for the
treatment and/or prevention of osteopenia, osteoporosis, secondary
osteoporosis, posttransplantation bone disease, Paget's disease,
idiopathic juvenile bone loss, multiple myeloma, osteosarcoma,
parathyroidectomy, thermal injuries, hyperparathyroidism,
periodontal diseases, bone disease caused by hemodyalysis, and
scurvy.
14. The pharmaceutical composition of claim 13, where secondary
osteoporosis is selected from the group consisting of
glucocorticoid induced osteoporosis, hyperthyroidisem induced
osteoporosis, immobilization induced osteoporosis, heparin induced
osteoporosis, and immunosuppressant induced osteoporosis.
15. The pharmaceutical composition of claim 11 or 12, for the
prevention and/or treatment of an abnormally low bone mineral
density (BMD).
16. A pharmaceutical composition for increasing BMD by decreasing
bone resorption comprising compound represented by the general
formula (I), ##STR00007## wherein R1, R2, and R3 represent i) a
hydrogen atom; or ii) a group having the formula CO--R wherein R is
a linear or branched alkyl group, saturated or unsaturated,
optionally substituted, and the main chain of said R contains from
1 to 25 carbon atoms; or iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer from 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group consisting of an oxygen atom, a sulphur atom, a
selenium atom, a CH.sub.2 group, a SO group, and a SO.sub.2 group;
iv) an entity selected from the group comprising
--PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R 2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a compound represented by the general formula
R''--COO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom,
a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer from 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; and R'' is a hydrogen atom or an alkyl group
containing from 1 to 4 carbon atoms; or a salt, prodrug or complex
of said compounds.
17. A pharmaceutical composition for increasing BMD by decreasing
bone resorption comprising compound represented by the general
formula (II), ##STR00008## wherein A1, A2 and A3 are chosen
independently and represent an oxygen atom, a sulphur atom or an
N-R4 group in which R4 is a hydrogen atom or a linear or branched
alkyl group, saturated or unsaturated, optionally substituted,
containing from 1 to 5 carbon atoms; wherein R1, R2, and R3
represent i) a hydrogen atom or a linear or branched alkyl group,
saturated or unsaturated, optionally substituted, containing from 1
to 23 carbon atoms; or ii) a group having the formula CO--R in
which R is a linear or branched alkyl group, saturated or
unsaturated, optionally substituted, and the main chain of said R
contains from 1 to 25 carbon atoms; or iii) a group having the
formula CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur
atom, a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group
or a SO.sub.2 group; n is an integer from 0 to 11; and R' is a
linear or branched alkyl group, saturated or unsaturated,
optionally substituted, wherein the main chain of said R' contains
from 13 to 23 carbon atoms and optionally one or more heterogroups
selected from the group consisting of an oxygen atom, a sulphur
atom, a selenium atom, a CH.sub.2 group, a SO group and a SO.sub.2
group; iv) an entity selected from the group comprising
--PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein RI, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a salt, prodrug or complex of said compound.
18. The pharmaceutical composition of claim 11, wherein the
compound is chosen from the group consisting of
tetradecylthioacetic acid (TTA), tetradecylselenoacetic acid, and
3-thia-15-heptadecyne.
19. The pharmaceutical composition of claim 16, wherein the
compound is chosen from the group consisting of
tetradecylthioacetic acid (TTA), tetradecylselenoacetic acid, and
3-thia-15-heptadecyne.
Description
THE FIELD OF THE INVENTION
[0001] Bone is a specialized dynamic connective tissue that serves
essential mechanical, protective, and metabolic functions. To
perform these functions efficiently bone must undergo a process
termed bone remodelling; continuous destruction (resorption) and
rebuilding (renewal) at millions of microscopic sites. During adult
life bone remodelling is crucial to eliminate structurally damaged
or aged bone and replace it with new healthy bone. To maintain the
proper bone mass resorption and formation must be kept in
equilibrium.
[0002] Various endogenous factors, such as age and diseases, as
well as exogenous influences such as injuries, drug treatments and
exposures may affect this equilibrium by increasing bone
resorption. The result can be observed as a decrease in the bone
mineral density (BMD), which measures the amount of calcium in the
bones that is closely correlated to total bone mass. Known
conditions that lead to a potentially pathological decrease in BMD
due to increased resorption are osteopenia, osteoporosis, secondary
osteoporosis, posttransplantation bone disease, Paget's disease,
idiopathic juvenile bone loss, multiple myeloma, osteosarcoma,
parathyroidectomy, thermal injuries, hyperparathyroidism,
periodontal diseases, bone disease caused by hemodyalysis and
scurvy.
[0003] With age the equilibrium between bone mass resorption and
formation becomes altered, generally in favour of resorption,
resulting in a reduction of bone mass termed osteopenia. This age
related loss of equilibrium is generally due to both a decline in
bone formation and more resorption. It is caused by the decrease in
estrogen production in post-menopausal women, and the decline with
age in the production of androgen in men (which is enzymatically
converted to estrogen, a hormone that regulates bone metabolism
directly and indirectly). Eventually this may lead to deterioration
of bone architecture, decreased resistance to stress, bone
fragility and susceptibility to fractures. These symptoms are
collectively referred to as osteoporosis, which is a major health
problem, especially in Western society, where it has been estimated
that up to 85% of women and somewhat fewer men older than 45 years
of age are at risk of developing osteoporosis.
[0004] In addition to age related osteoporosis, there are secondary
forms of osteoporosis not caused by age-related hormonal changes,
but by increased bone resorption due to exposure to
glucocorticoids, hyperthyroidisem, immobilizations, heparin and
immunosuppressants. The resulting conditions are generally referred
to as glucocorticoid induced osteoporosis, hyperthyroidisem induced
osteoporosis, immobilization induced osteoporosis, heparin induced
osteoporosis, and immunosuppressant induced osteoporosis.
[0005] There is a clear direct effect of glucocorticoids on bone
resorption in human cell systems, which explain the observed
increase in bone resorption seen in patients treated with
glucocorticoids. (Sivagurunathan S et al, J Bone Miner Res. 2005
March; 20(3):390-8. Epub 2004 Dec. 20)
[0006] Hyperthyroidism leads to secondary osteoporosis due to an
increase in bone resorption, and when it is treated, a prompt
decrease is found in bone resorption markers. (Inaba M, Clin
Calcium. 2001; 11(7):910-4)
[0007] During long term immobilizations, disuse induces dramatic
bone loss resulting from greatly elevated resorption. (Li C Y et
al, J Bone Miner Res. 2005 January; 20(1):117-24. Epub 2004 Oct.
18)
[0008] Osteoporosis is considered one of the potentially serious
side effects of heparin therapy. The pathogenesis is poorly
understood, but it has been suggested that heparin cause an
increase in bone resorption. (Gennari C et al, Aging (Milano).
June; 10(3):214-24)
[0009] Immunosuppressant therapy is known to lead to secondary
osteoporosis through increased bone resorption. (Kirino S et al, J
Bone Miner Metab. 2004; 22(6):554-60)
[0010] In association with transplantations large dosages of
immunosuppressive drugs are generally used. This leads to bone
resorption, which is referred to as posttransplantation bone
disease. (Cunningham J, Transplantation. 2005 Mar. 27;
79(6):629-34)
[0011] Paget's disease is not as common or as costly as
osteoporosis, but it affects 3% of the population over 40, and 10%
of the population over 80 years of age. Aside from causing bone
fractures it can lead to severe osteoarthritis and severe
neurological disorders. Paget's disease is characterized by rapid
bone turnover, caused by an increased bone resorption with
irregular bone formation, resulting in the formation of woven bone
of a tissue type formed initially in the embryo and during growth,
which is normally practically absent from the adult skeleton.
[0012] Idiopathic juvenile bone loss (osteoporosis) is a rare form
of bone demineralization that occurs during childhood. The
mechanism of bone loss is unclear, but some studies have found
increased bone resorption. (Bertelloni S et al, Calcif Tissue Int
1992 November; 51(5):400)
[0013] Multiple myeloma is a plasma cell malignancy characterized
by the high capacity to induce osteolytic bone lesions that mainly
result from an increased bone resorption related to the stimulation
of osteoclast recruitment and activity. (Giuliani N et al, Acta
Biomed Ateneo Parmense. 2004 December; 75(3):143-52)
[0014] Osteosarcoma leads to the destruction of bone tissue, and
genes for osteoclast differentiation have been found to be
differentially expressed in patients with osteosarcoma, thus
linking deviations in bone resorption to osteosarcoma. (Michelle B.
Mintz et al, Cancer Research. 2005 March:65, 1748-1754)
[0015] Routine hemodyalysis, as performed on patients with
extensive loss of retinal function, leads to an increase in bone
resorption, which causes bone diseases in this patient group.
(Hamano T et al, Bone. 2005 Mar. 24; Epub ahead of print)
[0016] Hyperparathyroidism is associated with skeletal changes
characterized by increased resorption and fibrous replacement of
bone. Parathyroidectomy in the treatment of secondary
hyperparathyroidism has also been shown to lead to an increase in
bone resorption, which may cause bone diseases in this patient
group. (Yajima A et al, Clin Calcium. 2003; 13(3):290-4)
[0017] In thermally injured children and adults there is a dramatic
decrease in bone formation which may be accompanied with an
increase in bone resorption. (Shea J E et al, J Musculoskelet
Neuronal Interact. 2003 September; 3(3):214-22)
[0018] Periodontal diseases are chronic infectious diseases that
result in loss of alveolar bone due to bone resorption triggered
through immune responses, and results from inflammatory reactions
directed against periodontopathic bacteria. (Ohmori Y, Clin
Calcium. 2001; 11(3):302-8)
[0019] Scurvy is caused by vitamin C depletion and leads to
structural collagen alterations, defective osteoid matrix formation
and increased bone resorption. Trabecular and cortical osteoporosis
is common in patients with scurvy.
[0020] Bone resorption is a specific function of osteoclasts, which
are multinucleated, specialized bone cells formed by the fusion of
mononuclear progenitors originating from the hemopoietic
compartment, more precisely from the granulocyte-macrophage
colony-forming unit (GM-CFU). The osteoclast is the principal cell
type to resorb bone, and together with the bone-forming cells, the
osteoblasts, dictate bone mass, bone shape and bone structure. The
increased activity and/or numbers of osteoclasts, relative to the
activity and or numbers of bone-forming osteoblasts, dictates the
development of osteoporosis and other diseases of bone loss.
[0021] For diseases in which osteoclasts presumably resorb bone at
abnormally high levels and osteoblasts form bone at normal levels,
the most reasonable therapeutic target for restoring the
equilibrium between bone resorption and formation would be
decreasing the number of osteoclasts and/or decreasing the
resorption activity of the osteoclasts. The treatments now
available for osteoporosis are indeed intended to suppress bone
resorption, but as their effects are variable and their side
effects may be substantial, there is room for improvement.
[0022] Osteoblasts are derived from bone marrow stromal cells. Upon
stimulation with glucocorticoids (GR) via the GR receptor, the bone
marrow stromal cells differentiate into osteoblasts. Upon
stimulation with peroxisome proliferator-activated receptor (PPAR)
ligands via the PPARs, the bone marrow stromal cells differentiate
into adiopocytes. The PPARs are members of the steroid nuclear
superfamily of receptors that are known as modulators of the
expression of genes involved in lipid metabolism and fat storage,
and there are three known types of PPARs; alpha, gamma and delta.
Based upon this, the "lipid hyphotehsis of osteoporosis" was
formed; the notion that osteoporosis may be caused by a high fat
diet upregulating adipocyte differentiation at the cost of
osteoblast differentiation. This hypotheses was later substantiated
by findings such as that oxidized lipids inhibit differentiation of
preosteoblasts, and that minimally oxidized LDL act to inhibit
osteogenic differentiation through activating PPAR alpha (Parhami F
et al, J Bone Miner Res, 1999 December; 14(12):2067-78), and that
PPAR gamma seems to be responsible for the decrease in osteoblasts
and increase in marrow fat seen in the elderly due to its
inhibition of osteoblastogenesis and stimulation of adipocyte
differentiation (Ali A A et al, Endocrinology. 2005 March;
146(3):1226-35. Epub 2004 Dec. 9), and that haploinnsufficency of
PPAR gamma promote osteogenesis through enhanced osteoblast
formation (Pei L J Clin Invest 2004 March; 113(6):805-6). Thus,
most of the prior art on osteoblasts and PPARs point to a role for
PPARs in inhibiting osteoblast differentiation, with the exception
of one publication which reported that activating PPAR alpha, delta
and gamma (with specific ligands) induced alkaline phosphatase
activity (which stimulate osteoblast maturation), matrix
calcification and the expression of osteoblast genes (Jackson S M
et al, FEBS Lett 2000 Apr. 7; 471(1):119-24). However, the same
publication also reported that at relatively high concentrations of
specific PPAR gamma ligands osteoblast maturation was inhibited.
Thus the prior art overall agrees with the lipid hyphotesis of
osteoporosis, indicating that stimulating the PPARs inhibit
osteoblast differentiation, thus decreasing bone formation.
[0023] Osteoclasts are derived from the monocyte-macrophage family.
Upon stimulation of the CFU-GM with macrophage colony stimulating
factor (M-CSF) form promonocytes which are immature nonadherent
progenitors of mononuclear phagocytes and osteoclasts. The
promonocytes may proliferate and differentiate along the macrophage
pathway, eventually forming a tissue macrophage, or may
differentiate along the osteoclast pathway, depending on the
cytokines to which they become exposed. Unlike for osteoblasts,
here is no obvious connection between osteoclast formation or
activity and PPARs, and very little research has been performed to
elucidate if such a connection exists. A PPAR gamma activator was
found to inhibit stimulators of osteoclastogenesis in one
publication, indirectly suggesting that PPAR gamma may decrease
bone resorption (Mbalaviele G, et al, J Biol. Chem. 2000 May 12;
275(19):14388-93), but a more recent publication found direct
evidence of the opposite; a PPAR gamma agonist enhanced bone loss,
increased fat marrow volume, and increased bone resorption
parameters (Sottile V et al Calcif Tissue Int. 2004 October;
75(4):329-37. Epub 2004 Jul. 13). Thus, nothing is known of PPAR
alpha and delta in regards to bone resorption, and the little
information there is about PPAR gamma is conflicting, although the
most direct evidence point towards a role for PPAR gamma in
increasing resorption.
[0024] In conclusion, the information available regarding the
overall effects of the PPARs on the equilibrium between bone mass
resorption and formation is very sketchy, but points toward PPAR
agonists having a negative effect on the BMD, both through
decreased bone formation and increased bone resorption.
[0025] Thus, when the inventor found that the PPAR alpha, gamma and
delta agonist tetradecylthioacetic acid (TTA) stimulated BMD
increase and inhibited bone resorption, this was quite
unexpected.
[0026] The use of compounds according to the invention is thus
characterized by that the compound is represented by the general
formula (I),
##STR00001##
wherein R1, R2, and R3 represent [0027] i) a hydrogen atom; or
[0028] ii) a group having the formula CO--R in which R is a linear
or branched alkyl group, saturated or unsaturated, optionally
substituted, and the main chain of said R contains from 1 to 25
carbon atoms; or [0029] iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; [0030] iv) an entity selected from the group
comprising --PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a compound represented by the general formula
R''--COO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom,
a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; and R'' is a hydrogen atom or an alkyl group
containing from 1 to 4 carbon atoms; or a salt, prodrug or complex
of said compounds, or the compound is represented by the general
formula (II),
##STR00002##
[0030] wherein A1, A2 and A3 are chosen independently and represent
an oxygen atom, a sulphur atom or an N-R4 group in which R4 is a
hydrogen atom or a linear or branched alkyl group, saturated or
unsaturated, optionally substituted, containing from 1 to 5 carbon
atoms; wherein R1, R2, and R3 represent [0031] i) a hydrogen atom
or a linear or branched alkyl group, saturated or unsaturated,
optionally substituted, containing from 1 to 23 carbon atoms; or
[0032] ii) a group having the formula CO--R in which R is a linear
or branched alkyl group, saturated or unsaturated, optionally
substituted, and the main chain of said R contains from 1 to 25
carbon atoms; or [0033] iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; [0034] iv) an entity selected from the group
comprising --PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a salt, prodrug or complex of said compound, for the
preparation of a pharmaceutical composition for the prevention
and/or treatment of conditions associated with low/decreased bone
mineral density (BMD), and/or for increasing the BMD by decreasing
the bone resorption.
DETAILED DESCRIPTION OF THE INVENTION
[0035] TTA is a non .beta.-oxidizable fatty acid analogue,
belonging to a group of compounds comprising
a compound represented by the general formula (I),
##STR00003##
wherein R1, R2, and R3 represent [0036] i) a hydrogen atom; or
[0037] ii) a group having the formula CO--R in which R is a linear
or branched alkyl group, saturated or unsaturated, optionally
substituted, and the main chain of said R contains from 1 to 25
carbon atoms; or [0038] iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; [0039] iv) an entity selected from the group
comprising --PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a compound represented by the general formula
R''--COO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom,
a selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; and R'' is a hydrogen atom or an alkyl group
containing from 1 to 4 carbon atoms; or a salt, prodrug or complex
of said compounds; or a compound represented by the general formula
(II),
##STR00004##
[0039] wherein A1, A2 and A3 are chosen independently and represent
an oxygen atom, a sulphur atom or an N-R4 group in which R4 is a
hydrogen atom or a linear or branched alkyl group, saturated or
unsaturated, optionally substituted, containing from 1 to 5 carbon
atoms; wherein R1, R2, and R3 represent [0040] i) a hydrogen atom
or a linear or branched alkyl group, saturated or unsaturated,
optionally substituted, containing from 1 to 23 carbon atoms; or
[0041] ii) a group having the formula CO--R in which R is a linear
or branched alkyl group, saturated or unsaturated, optionally
substituted, and the main chain of said R contains from 1 to 25
carbon atoms; or [0042] iii) a group having the formula
CO--(CH.sub.2).sub.2n+1--X--R', wherein X is a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group or a
SO.sub.2 group; n is an integer of 0 to 11; and R' is a linear or
branched alkyl group, saturated or unsaturated, optionally
substituted, wherein the main chain of said R' contains from 13 to
23 carbon atoms and optionally one or more heterogroups selected
from the group comprising an oxygen atom, a sulphur atom, a
selenium atom, an oxygen atom, a CH.sub.2 group, a SO group and a
SO.sub.2 group; [0043] iv) an entity selected from the group
comprising --PO.sub.3CH.sub.2CHNH.sub.3COOH (serine),
PO.sub.3CH.sub.2CH.sub.2NH.sub.3 (ethanolamine),
PO.sub.3CH.sub.2CH.sub.2N(CH.sub.3).sub.3 (choline),
PO.sub.3CH.sub.2CHOHCH.sub.2OH (glycerol) and PO.sub.3(CHOH).sub.6
(inositol); wherein R1, R2, and R3 are chosen independently from
i), ii), iii), or iv), but at least one of R1, R2, or R3 is defined
by iii); or a salt, prodrug or complex of said compound.
[0044] In a preferred embodiment of a compound according to the
invention at least one of R1, R2 or R3 is an alkyl.
[0045] In a preferred embodiment of a compound according to the
invention at least one of R1, R2 or R3 is an alkene.
[0046] In a preferred embodiment of a compound according to the
invention at least one of R1, R2 or R3 is an alkyne.
[0047] In a preferred embodiment of a compound according to the
invention at least one of R1, R2 or R3 is tetradecylthioacetic
acid.
[0048] In a preferred embodiment of a compound according to the
invention at least one of R1, R2 or R3 is tetradecylselenoacetic
acid.
[0049] Preferred embodiments of the compounds according to the
invention are tetradecylthioacetic acid (TTA),
tetradecylselenoacetic acid and 3-Thia-15-heptadecyne.
[0050] In a preferred embodiment of a compound according to the
invention n is 0 or 1.
[0051] In a preferred embodiment of a compound according to the
invention said compound is a phospholipid, wherein said
phospholipid is selected from the group comprising phosphatidyl
serine, phosphatidyl choline, phosphatidyl ethanolamine,
phosphatidyl inositol, phosphatidyl glycerol, diphosphatidyl
glycerol.
[0052] In a preferred embodiment of a compound according to the
invention said compound is a triacylglycerol.
[0053] In a preferred embodiment of a compound according to the
invention said compound is a diacylglycerol.
[0054] In a preferred embodiment of a compound according to the
invention said compound is a monoacylglycerol.
[0055] In a preferred embodiment of a compound according to formula
(II) A1 and A3 both represent an oxygen atom, while A2 represent a
sulphur atom or an N-R4 group in which R4 is a hydrogen atom or a
linear or branched alkyl group, saturated or unsaturated,
optionally substituted, containing from 1 to 5 carbon atoms.
[0056] The compounds according to the invention are analogues of
naturally occurring compounds, and as such are recognized by the
same systems which process the natural compounds, including the
enzymes that .beta.- and in some cases .omega.-oxidize natural long
chain fatty acids. The analogues differ from their naturally
occurring counterparts in that they cannot be completely oxidized
in this manner.
[0057] The compounds according to the invention may be non
.beta.-oxidizable fatty acid analogues, as represented by the
formula R''CCO--(CH.sub.2).sub.2n+1--X--R'. However, said compounds
may also be more complex structures derived from one or more of
said non .beta.-oxidizable fatty acid analogues, as represented by
the general formulas (I) or (II). These compounds are analogues of
naturally occurring mono-, di-, and triacylglycerols, or
phospholipids including phosphatidyl serine, phosphatidyl choline,
phosphatidyl ethanolamine, phosphatidyl inositol, phosphatidyl
glycerol, and diphosphatidyl glycerol. Said compounds may also
comprise a substitution in the glycerol backbone, as shown in
formula (II). Said substitution of the oxygen(s) is achieved by
replacing the oxygen(s) with sulphur or a nitrogen containing
group. This may block hydrolysis before uptake by the intestines,
thus increasing the bioavailability of the compounds.
[0058] The above complex structures derived from one or more of
said non .beta.-oxidizable fatty acid analogues have their effect
because the fatty acid analogues they comprise are not capable of
being fully .beta.-oxidized. Said complex structures may have an
effect as complete structures, and as naturally resulting
degradation products comprising the fatty acid analogues. Because
the compounds are not able to be fully .beta.-oxidized, they will
build up, and this triggers an increase in the .beta.-oxidation of
naturally occurring fatty acids. Many of the effects of the
compounds according to the invention are due to this increase in
.beta.-oxidation.
[0059] During .beta.-oxidation, a fatty acid is enzymatically
oxidized cleaved between carbons 2 and 3 (when counting from the
carboxylic end of the fatty acid), resulting in the removal of the
two carbon atoms on either side of the oxidation site as acetic
acid. This step is then repeated on the now two carbons shorter
fatty acid, and repeated again until the fatty acid is fully
oxidized. .beta.-oxidation is the usual way in which the majority
of fatty acids are catabolized in vivo. The .beta.-oxidation
blocking by the compounds according to the invention is achieved by
the insertion of a non-oxidizable group in the X position in the
formula of the present invention. Because the mechanism for
.beta.-oxidation is well known, X is defined as S, O, SO, SO.sub.2,
CH.sub.2 or Se. Anyone skilled in the art would assume, without an
inventive step, that these compounds would all block
.beta.-oxidation in the same manner.
[0060] In addition, the compounds may contain more than one block,
i.e. in addition to X, R' may optionally comprise one or more
heterogroups selected from the group comprising an oxygen atom, a
sulphur atom, a selenium atom, an oxygen atom, a CH.sub.2 group, a
SO group and a SO.sub.2 group. As an example, one may insert two or
three sulphurs as X to induce a change in the degradation of the
fatty acid and thus a modulated effect. Multiple sulphur atoms
would also modulate the polarity and stability somewhat. From a
pharmacological viewpoint it is generally desirable to be able to
present a spectrum of compounds rather than just one single
compound to avoid or counteract problems with resistance.
[0061] In addition to the identity of X, its position is also an
issue. The distance of X from the carboxylic end of the fatty acid
is defined by how many CH.sub.2 groups are positioned between X and
the carboxylic end of the fatty acid, which is defined by
(CH.sub.2).sub.2n+1, where n is an integer of 0 to 11. Thus there
are an odd number of CH.sub.2 groups, that is; the position of X
relative to the carboxyl group is such that X eventually blocks
.beta.-oxidation. The range of n is chosen to include all
variations of the fatty acid analogue which has the desired
biological effect. Since .beta.-oxidation in theory can work on
infinitely long molecules, n could be infinite, but in practice
this is not so. The fatty acids which normally undergo
.beta.-oxidation are usually 14 to 24 carbon atoms long, and this
length is therefore most ideal for undergoing enzymatic
.beta.-oxidation. The ranges of n and R' are thus given so that the
fatty acid analogues will cover this range. (Likewise, option ii)
of formulas (I) and (II) and define R to have 1 to 25 carbon
groups, and option i) of formula (II) define the alkyl group to
contain from 1 to 23 carbon atoms, to be analogous to naturally
occurring compounds.) The total number of carbon atoms in the fatty
acid backbone is preferably between 8 and 30, most preferably
between 12 and 26. This size range is also desirable for the uptake
and transport through cell membranes of the fatty acid analogues of
the present invention.
[0062] Although all fatty acid anagoges with an odd positioning of
the .beta.-oxidation blocker X away from the carboxylic end block
.beta.-oxidation, the extent of their biological effect may be
variable. This is due to the difference in biological degradation
time of the various compounds. The inventors have done experiments
to show the effect of moving X further from the carboxylic fatty
acid end. In these experiments the activity (in
nmol/min/mg/protein) of mitochondrial .beta.-oxidation in the liver
of fatty acid analogues was measured with sulphur in the 3, 5 and 7
positions relative to the carboxyl end. The activities were 0.81
for sulphur in the 3.sup.rd position, 0.61 for sulphur in the
5.sup.th position, 0.58 for sulphur in the 7.sup.th position, and
0.47 for palmitic acid, the non .beta.-oxidation blocking control.
This shows, as expected, that .beta.-oxidation is indeed blocked by
fatty acid analogues with varying positioning of the block, and
that the effect thereof is lessened the further away from the
carboxylic end the block is positioned at, because it takes the
.beta.-oxidation longer to reach the block so more of the fatty
acid analogue is degraded by then. However, as the decline is great
for going from the 3.sup.rd to 5.sup.th position, but small going
from the 5.sup.th to 7.sup.th position, it is reasonable to assume
that this decline will continue to be less as one moves out the
chain, and thus that it will be very far out indeed before no
effect (compared to the control) is seen at all.
[0063] Thus, it is reasonable to include as compounds of the
present invention, fatty acid analogues and other compounds
represented by the general formulas (I) and (II), (which comprise
said fatty acid analogue(s),) which block .beta.-oxidation at
different distances from the carboxylic end of the analogues, as
the compounds of the present invention all do indeed block
.beta.-oxidation, even if the effect thereof can be modulated. This
modulation will after all differ under wearying conditions; in
different tissues, with wearying dosages, and by changing the fatty
acid analogue so that it is not so easily broken down, as will be
described next. Thus it is reasonable to include in the formula all
distances of the .beta.-oxidation blocker from the carboxylic end
of the fatty acid analogue which are biologically relevant.
[0064] Although fatty acid analogues as described with a block in
the X position cannot undergo .beta.-oxidation, they may still
undergo .omega.-oxidation. This is a much less common and slower
biological process, which oxidizes the fatty acid not from the
carboxylic end, but rather from the methyl/hydrophobic head group,
here termed R'. In this pathway the carbon atom at the .omega.-end
of the fatty acid is hydroxylated by a member of the cytochrome
P450 enzyme family. This hydroxylated fatty acid is then converted
into an aldehyde by an alcohol dehydrogenase, and subsequently this
aldehyde is converted into a carboxyl group by an aldehyde
dehydrogenase. As a consequence, the final product of the pathway
is a dicarboxylic fatty acid, which can be degraded further by
.omega.-oxidation from the .omega.-end.
[0065] .omega.-oxidation is believed to be the main pathway for
degradation of the fatty acid analogues as described with a block
in the X position. Experiments were thus performed where R' was
changed to block w-oxidation, by introducing a triple bond at the
methyl end of the fatty acid analogue. This resulted in the fatty
acid analogue 3-thia-15-heptadecyn, which when tested showed the
expected result: a substantially increased degradation time in
vivo. This is important for the use of the fatty acid analogues in
pharmaceutical preparation, as it may potentiate the effects of the
.beta.-oxidizable fatty acid analogues by further slowing down
their breakdown.
[0066] Again, as with the blocking of .beta.-oxidation, it is
routine to find other fatty acid analogues witch would block
.omega.-oxidation in exactly the same manner, based upon knowledge
of how .omega.-oxidation occurs. A double bond will for instance
have the exact same effect as the triple bond did, and it is
therefore included in the definition of the methyl/hydrophobic head
group end of the molecule, here termed R', that it may be saturated
or unsaturated. A branch may also block oxidation, so R' is defined
as linear or branched.
[0067] In order to block .omega.-oxidation by the insertion of a
substitute in R', said R' may be substituted in one or several
positions with heterogroups selected from the group comprising an
oxygen atom, a sulphur atom, a selenium atom, an oxygen atom, a
CH.sub.2 group, a SO group and a SO.sub.2 group. R' may also be
substituted with one or more compounds selected from the group
comprising fluoride, chloride, hydroxy, C.sub.1-C.sub.4 alkoxy,
C.sub.1-C.sub.4 alkylthio, C.sub.2-C.sub.5 acyloxy or
C.sub.1-C.sub.4 alkyl.
[0068] Thus the compounds according to the present invention are
either fatty acids analogous to naturally occurring fatty acids,
which are not capable of being .beta.-oxidized, or naturally
occurring lipids comprising said fatty acid analogues. In vivo, the
fatty acid analogues show a strong preference for being
incorporated into phospholipids. In some cases it is indeed
advantageous to mimic nature and incorporate the fatty acid
analogues in naturally occurring lipids, such as mono-, di-, and
triglycerides and phospholipids. This changes the absorption of the
compounds (when comparing fatty acids to fatty acids incorporated
in larger lipid structures) and may increase the bioavailability or
stability.
[0069] As an example, one could make a complex by including a fatty
acid(s) which are not capable of being .beta.-oxidized into a
triacylglycerol. Such compounds are encompassed by formulas (I) and
(II). If such a triacylglycerol was taken orally, for instance in
an animal feed product, it would probably be transported like any
triacylglycerol, from the small intestine in chylomicrons and from
the liver in the blood in lipoproteins to be stored in the adipose
tissue or used by muscles, heart or the liver, by hydrolyzes of the
triacylglycerol into glycerol and 3 free fatty acids. The free
fatty acids would at this point be the parent compound of the
present invention, and not a complex anymore.
[0070] Yet other possible glycerophospholipid derivatives of the
fatty acids of the present invention includes, but are not limited
to, phosphatidyl cholines, phosphatidyl ethanolamines, phosphatidyl
inositols, phosphatidyl serines and phosphatidyl glycerols.
[0071] Another esterification of fatty acids found in vivo which
could be easily used to make a complex for a compound of the
present invention would be to make the alcohol or polyalcohol
corresponding to the fatty acid, for example one could make a
sphingolipid derivative such as ceramide or sphingomyelin by making
the corresponding amino alcohol. Like the glycerophospholipid
complexes, such complexes would be very water insoluble and less
hydrophilic. These kinds of hydrophobic complexes of the present
invention would pass easier through biological membranes.
[0072] Other possibilities of polar complexes of the present
invention may be, but are not limited to, lysophospholipids,
phosphatidic acis, alkoxy compounds, glycerocarbohydrates,
gangliosiedes, and cerebrosides.
[0073] Although there can be large structural differences between
different compounds of the invention, the biological functions of
all compounds are expected to be similar because they all block
.beta.-oxidation in the same manner. This inability of the lipid
analogues to be .beta.-oxidized (and in some cases,
.omega.-oxidized,) causes the analogues to build up in the
mitochondria, which triggers the .beta.-oxidation of the in vivo
naturally occurring fatty acids, which in turn leads to many of the
biological effects of the fatty acid analogues of the present
invention. (Berge R K et al. (2002) Curr Opin Lipidol
13(3):295-304)
[0074] The peroxisome proliferator-activated receptor (PPAR) family
are pleiotropic regulators of cellular functions such as cellular
proliferation, differentiation and lipid homeostasis (Ye J M et al.
(2001) Diabetes 50:411-417). The PPAR family is comprised of three
subtypes; PPAR.alpha., PPAR.beta., and PPAR.gamma.. A fatty acid
analogue according to formula (I); TTA, has been used previously by
the present inventors to test the various biological effects of the
fatty acid analogues. TTA is a potent ligand of PPAR.alpha. (Forman
B M, Chen J, Evans R M (1997) Proc Natl Acad Sci 94:4312-4317;
Gottlicher M et al. (1993) Biochem Pharmacol 46:2177-2184; Berge R
K et al. (1999) Biochem J 343(1):191-197). As a PPAR.alpha.
activator TTA stimulate the catabolism of fatty acids by increasing
their cellular uptake. Lowering the plasma triglyceride levels with
TTA caused a shift in liver cellular metabolism, towards
PPAR.alpha. regulated fatty acid catabolism in mitochondria. (Grav
H J et al. (2003) J Biol Chem 278(33):30525-33). TTA also activate
PPAR.beta. and PPAR.gamma. as well as PPAR.alpha. (Raspe E et al.
(1999) J Lipid Res 40:2099-2110). It is believed that the fatty
acid analogues according to formula (I) will have the same effects
as PPAR activators as exemplified by TTA, since their biological
effects on fatty acid oxidation is the same, and they are believed
to be PPAR activators due to their ability to block fatty acid
oxidation.
[0075] In the current invention, TTAs effect on bone resorption in
vivo and bone mineral density (BMD) in rats was investigated. In
addition, the effects on these parameters of the PPAR.alpha.
agonists Wyeth 14,643 and fenofibrate, and the PPAR.gamma. agonist
pioglitazone was also investigated.
[0076] The in vivo experiments on rats showed an increase in
femoral BMD for rats treated with PPAR.alpha. agonists Wyeth 14,643
and fenofibrate, of 7% and 5.7% respectively. As discussed, there
was little prior art available to suggest what one should have
expected for PPAR.alpha. agonists. The PPAR.gamma. agonist
pioglitazone induced a decrease in femoral BMD of -3%, which is as
expected from the prior art. TTA is both a PPAR.alpha. and
PPAR.gamma. agonist, so one would most probably assume that the
result of it being a PPAR.alpha. agonist was unclear, while the
result of it being a PPAR.gamma. agonist would be a reduction in
BMD, and the overall effect of TTA was thus not very predictable,
but probably leaning towards a reduction in BMD. TTA did however
show the unexpected result of increasing the femoral BMD by 5%.
[0077] The in vitro experiments on preosteoblasts showed similar
results for resorption. PPAR.alpha. agonists Wyeth 14,643 and
fenofibrate, as well as PPAR.alpha. and PPAR.gamma. agonist TTA,
stimulated OPG and interlukin 6 (IL-6) release from preosteoblasts,
while the PPAR.gamma. agonist pioglitazone did the opposite. OPG is
an important inhibitor of bone resorption, while IL-6 is an
important stimulator of bone resorption. Thus, the overall effect
is that of bone resorption inhibition by Wyeth 14,643, fenofibrate
and TTA, and bone resorption stimulation by pioglitazone.
[0078] Seeing as an increase in BMD may be caused by an increase in
bone formation or a decrease in bone resorption, the increase in
BMD for Wyeth 14,643, fenofibrate and TTA may be due to the
decrease in bone resorption, and the decrease in BMD for
pioglitazone may be due to the increase in bone resorption.
However, the changes in bone resorption may also only be partially
responsible for the changes in BMD; there may also be a concurrent
change in bone formation.
[0079] The BMD is closely correlated to total bone mass, and loss
thereof is associated with a host of diseases. Loss of bone mass
due to an increase in resorption compared to that seen in healthy
individuals is also associated with diseases, as discussed earlier.
By increasing the BMD and decreasing the resorption TTA and the
other fatty acid analogues according to the present invention are
therefore potentially beneficial in treating or preventing these
diseases associated with a loss in BMD and/or increase in bone
resorption.
Administration of the Compounds of the Present Invention
[0080] As a pharmaceutical medicament the compounds of the present
invention may be administered directly to the animal by any
suitable technique, including parenterally, intranasally, orally,
or by absorption through the skin. They can be administered locally
or systemically. The specific route of administration of each agent
will depend, e.g., on the medical history of the recipient human or
animal.
[0081] Examples of parenteral administration include subcutaneous,
intramuscular, intravenous, intra-arterial, and intra-peritoneal
administration
[0082] As a general proposition, the total pharmaceutically
effective amount of each of the non .beta.-oxidizable fatty acid
analogues administered parenterally per dose will preferably be in
the range of about 1 mg/kg/day to 200 mg/kg/day of patient body
weight for humans, although, as noted above, this will be subject
to a great deal of therapeutic discretion. A dose of 5-50 mg/kg/day
is most preferable.
[0083] If given continuously, the compounds of the present
invention are each typically administered by 1-4 injections per day
or by continuous subcutaneous infusions, for example, using a
mini-pump. An intravenous bag solution may also be employed.
[0084] For parenteral administration, in one embodiment, the
compounds of the present invention are formulated generally by
mixing each at the desired degree of purity, in a unit dosage
injectable form (solution, suspension, or emulsion), with a
pharmaceutically acceptable carrier, i.e., one that is non-toxic to
recipients at the dosages and concentrations employed and is
compatible with other ingredients of the formulation.
[0085] Generally, the formulations are prepared by contacting the
compounds of the present invention each uniformly and intimately
with liquid carriers or finely divided solid carriers or both.
Then, if necessary, the product is shaped into the desired
formulation. Preferably the carrier is a parenteral carrier, more
preferably a solution that is isotonic with the blood of the
recipient. Examples of such carrier vehicles include water, saline,
Ringer's solution, and dextrose solution. Non-aqueous vehicles such
as fixed oils and ethyl oleate are also useful herein, as well as
liposomes.
[0086] The carrier may suitably contain minor amounts of additives
such as substances that enhance isotonicity and chemical stability.
Such materials are non-toxic to recipients at the dosages and
concentrations employed, and include buffers such as phosphate,
citrate, succinate, acetic acid, and other organic acids or their
salts; antioxidants such as ascorbic acid; immunoglobulins;
hydrophilic polymers such as polyvinylpyrrolidone; amino acids,
such as glycine, glutamic acid, aspartic acid, or arginine;
monosaccharides, disaccharides, and other carbohydrates including
cellulose or its derivatives, glucose, mannose, or dextrins;
chelating agents such as EDTA; sugar alcohols such as mannitol or
sorbitol; counterions such as sodium; and/or non-ionic surfactants
such as polysorbates, poloxamers, or PEG.
[0087] For oral pharmacological compositions such carrier material
as, for example, water, gelatine, gums, lactose, starches,
magnesium-stearate, talc, oils, polyalkene glycol, petroleum jelly
and the like may be used. Such pharmaceutical preparation may be in
unit dosage form and may additionally contain other therapeutically
valuable substances or conventional pharmaceutical adjuvants such
as preservatives, stabilising agents, emulsifiers, buffers and the
like. The pharmaceutical preparations may be in conventional liquid
forms such as tablets, capsules, dragees, ampoules and the like, in
conventional dosage forms, such as dry ampoules, and as
suppositories and the like.
[0088] In addition the compounds of the present invention, i.e. the
fatty acid analogue according to formula (I), may be used in
nutritional preparations, as defined earlier, in which case the
dosage of the fatty acid analogue preferable is as described
pharmaceuticals or less. In animal fodder, the amount of non
.beta.-oxidizable fatty acid analogue can be up to 10 times that in
products for human consumption, that is, up to 2 g/kg/day of animal
body weight.
Measuring Bone Parameters
[0089] In many cases it is obvious whether the use of fatty acid
analogues in accordance with the present invention is appropriate
or not, and this choice can be made by the skilled medical
professional without undue experimentation. If a person is for
instance about to undergo aggressive immunosuppressant therapy, it
would be routine for the skilled medical professional to determine
if bone loss could be prevented by the use of compounds according
to formula (I). However, there may be instances where it is
necessary to first determine if bone metabolism impairment has
occurred, or to determine the bone density, or to determine the
boner resorption.
[0090] There are several techniques currently used by medical
professionals to measure bone metabolism impairment. One can
perform a bone cell density measurement, or an ultrasound
densitometer test, or measure bone metabolism parameters directly.
Bone metabolism parameters comprise markers present in the blood
such as BAP (a marker of osteogenesis), DPD (a bone resorption
marker), NTX, corrected Ca, and the SOS value (an index of bone
metabolism impairment).
[0091] Several methods are available to measure bone density, but
currently the most widely used technique is DEXA (Dual Energy Xray
Absorptiometry). This is the method used to determine efficacy in
the recent large clinical trials, and to characterize fracture risk
in large epidemiological studies. Older methods such as single
photon absorptiometry do not predict hip fractures as well as DEXA.
Three companies manufacture these densitometers: Hologic, Norland,
and Lunar.
[0092] Newer techniques such as ultrasound appear to offer a more
cost-effective method of screening bone mass. Ultrasound
measurements are usually performed at the calcaneous and it is not
possible to measure sites of osteoporotic fracture such as the hip
or spine. Adding an ultrasound measurement to a DEXA does not
improve the prediction of fractures.
[0093] Quantitative computed tomography (QCT) is another method for
measuring bone density. QCT of the spine must be done following
strict protocols in laboratories that do these tests frequently; in
community settings the reproducibility is poor. The QCT
measurements decrease more rapidly with aging, so the "T scores" in
older individuals will be much lower than DEXA measurements.
[0094] Several techniques can measure bone density at the hand,
radius or ankle. These include single energy absorptiometry,
metacarpal width or density from hand xrays. Magnetic resonance
imaging is a new method of measuring bone density.
[0095] Measuring the bone density gives a good indication of how
healthy the bones are. However, if it is not as one would desire, a
bone density measurement in itself does not provide information
regarding on whether there this is due to too much resorption or
too little new bone formation, and to find out one must measure
bone resorption and/or formation directly.
[0096] Bone resorption markers are released in circulation as
byproducts of osteoclast action on bone and include cross-links for
collagen type I. Bone formation markers are released during
osteoblast synthesis of new bone protein matrix, which can be
assessed by measuring circulating osteocalcin.
[0097] Bone resorption can be measured by the Crosslaps method.
Crosslaps is degradation products of C-terminal telopeptides of
type I collagen in human serum and plasma, which can be measured in
urine with specific RIA and ELISAs. Since more than 90% of the
organic matrix of bone consists of type 1 collagen, measuring its
degradation products in urine makes crosslaps a potential specific
marker of bone resorption. A pronounced and significant increase
(47-142%) in Crosslaps at menopause indicate that it is a very
sensitive marker of metabolic bone changes taking place at
menopause. The correlation between Crosslaps and the rate of loss
measured by single photon absorptiometry has been shown to be much
higher than with Pyridinoline and Deoxypyridinoline. Crosslaps has
also been used to predict the rate of bone loss considering a bone
loss of more than 3% per year. Crosslapse has a specificity of 80%
and a sensitivity of more than 70%, it can thus be used as a
potentially useful screening parameter in the risk assessment of
diseases like postmenopausal osteoporosis and Paget's disease.
Crosslaps values decreases substantially in response to replacement
therapies thus suggesting its usefulness in monitoring treatment
efficacy. Crosslaps have been cleared by the FDA, and is currently
used for research only, but is expected to be available for general
testing soon. It is developed by Nordic bioscience diagnostics.
[0098] Another method for measuring bone resorption which is
currently used clinically is measuring pyridinoline and
deoxypyridinoline (Pyr and D-Pyr). Pyridinoline cross links are
released into the circulation during bone resorption and are
excreted as both free and bound to C and N terminal ends of type 1
collagen. These are measured by different methods, the most
sensitive being HPLC. More recently sensitive chemiluminescence
based assays have also been made available.
[0099] Fasting urinary calcium measured in a morning sample and
corrected for creatinine excretion is the cheapest marker of bone
resorption. It is useful to detect marked changes in bone
resorption but lacks sensitivity especially in conditions
characterized by subtle alteration of bone turnover such as
osteoporosis. Hydroxyproline is found mainly in collagen and
represents about 13% of the amino acid content of the molecule.
Hydroxyproline is highly metabolized before being excreted and is
poorly correlated with bone resorption as assessed by calcium
kinetic and bone histomorphometry.
Experimental SectionEXPERIMENTAL SECTION
[0100] The preparation of non .beta.-oxidizable fatty acid
analogues according to the present invention is disclosed in detail
in the applicant's earlier Norwegian patent applications no.
20005461, 20005462, 20005463 and 20024114. These documents also
describe toxicity studies of TTA. Preparation of mono-, di-, and
triglycerides and nitrogen comprising lipids according to the
invention is disclosed in detail in U.S. patent application Ser.
No. 10/484,350. The preparation of phospholipids including serine,
ethanolamine, choline, glycerol, and inositol according to the
invention is disclosed in detail in the applicant's earlier
Norwegian patent application no. 20045562.
Example 1
Biological Effects of the Composition According to the Invention:
In Vitro Studies of Bone Resorption
[0101] The effect of the PPAR agonists on release of
osteoprotegerin (OPG), RANKL and IL6 from the preosteoblast cell
line MC3T3-E1 was investigated. Wyeth 14,643 was found to inhibit
the differentiation of human monocytes into osteoclasts in a
dose-dependent manner, and also inhibited the proliferation of the
preosteoclast cell line RAW264.7. Wyeth 14643, fenofibrate and TTA
were also shown to stimulate OPG release and inhibit IL6 release
from preosteoblasts, while no effects on RANKL could be detected.
Pioglitazone, on the other hand, tended to inhibit the OPG release
and stimulated the release of IL6.
Example 2
Biological Effects of the Composition According to the Invention:
In Vivo Studies of BMD
[0102] Fifty female Fischer rats were divided into 5 groups and
were given methocel (control group), Wyeth 14,643, fenofibrate,
tetradecylthioacetic acid (TTA) and pioglitazone (50 mg/kg body
weight) by intragastric gavage for 4 months. Body weight was
registered throughout the study. BMD was measured by double x-ray
absorptiometry (DXA). Histomorhometry of femur was performed, and
mechanical strength in the femoral shaft and collum femoris was
measured.
[0103] There was no difference in body weight between control rats
and the treatment groups at the end of the study. After 4 months
femoral BMD was significantly higher in rats treated with Wyeth
14,643 (7%), fenofibrate (5.7%) and TTA (5%) than in control rats.
In rats treated with pioglitazone, femoral BMD tended to be lower
than in control rats (-3%).
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