U.S. patent application number 10/154625 was filed with the patent office on 2003-01-16 for methods for selectively occluding blood supplies to neoplasias.
This patent application is currently assigned to EFA Sciences. Invention is credited to Das, Undurti N..
Application Number | 20030013759 10/154625 |
Document ID | / |
Family ID | 25483994 |
Filed Date | 2003-01-16 |
United States Patent
Application |
20030013759 |
Kind Code |
A1 |
Das, Undurti N. |
January 16, 2003 |
Methods for selectively occluding blood supplies to neoplasias
Abstract
Disclosed are methods of selectively reducing the blood supply
to a neoplastic region, such as a tumor region, thereby selectively
causing necrosis of the neoplastic tissue without substantial
necrosis of adjoining tissues. In particular, methods are disclosed
of selectively reducing the blood supply to a neoplastic region,
such as a tumor region, by causing selectively occlusion of blood
vessels feeding the neoplastic region. The invention also provides
methods of selectively causing anti-angiogenic action in a
neoplastic region, such as a tumor region, with the result that new
blood vessels are not formed to sustain the neoplasia. The methods
employ intra-arterial injection of polyunsaturated fatty acids,
preferably in the form of salts, preferably with a lymphographic
agent, and optionally with an anti-cancer drug, and/or a cytokine.
The invention also provides solutions of PUFAs, or salts of PUFAs,
in combination with a lymphographic agent.
Inventors: |
Das, Undurti N.; (Norwood,
MA) |
Correspondence
Address: |
HALE AND DORR, LLP
60 STATE STREET
BOSTON
MA
02109
|
Assignee: |
EFA Sciences
|
Family ID: |
25483994 |
Appl. No.: |
10/154625 |
Filed: |
May 24, 2002 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10154625 |
May 24, 2002 |
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09946129 |
Sep 4, 2001 |
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6426367 |
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09946129 |
Sep 4, 2001 |
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09392953 |
Sep 9, 1999 |
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Current U.S.
Class: |
514/492 ;
514/494; 514/499; 514/560 |
Current CPC
Class: |
A61K 31/28 20130101;
A61K 33/00 20130101; A61K 33/00 20130101; A61K 31/28 20130101; A61P
35/00 20180101; A61K 31/201 20130101; A61K 49/0461 20130101; A61K
31/202 20130101; A61K 45/06 20130101; A61K 31/202 20130101; A61K
31/557 20130101; A61K 2300/00 20130101; A61K 31/201 20130101; A61K
2300/00 20130101; A61K 2300/00 20130101; A61K 49/0476 20130101;
A61K 2300/00 20130101 |
Class at
Publication: |
514/492 ;
514/494; 514/499; 514/560 |
International
Class: |
A61K 031/315; A61K
031/28; A61K 031/202 |
Claims
What is claimed is:
1. A pharmaceutical composition comprising a polyunsaturated fatty
acid, or a salt of a polyunsaturated fatty acid, in combination
with a lymphographic agent.
2. A pharmaceutical composition as in claim 1 wherein said
lymphographic agent is an iodized fatty acid.
3. A pharmaceutical composition as in claim 1 wherein said
lymphographic agent is combined with said polyunsaturated fatty
acid in a solution.
4. A pharmaceutical composition as in claim 1 wherein said
lymphographic agent is conjugated to said polyunsaturated fatty
acid.
5. A pharmaceutical composition as in any one of claims 1-4 wherein
said polyunsaturated fatty acid is an essential fatty acid.
6. A pharmaceutical composition as in claim 5 wherein said
essential fatty acid is selected from the group consisting of
gamma-linolenic acid, arachidonic acid, docosahexaenoic acid,
eicosapentaenoic acid, di-homo-gamma-linolenic acid,
alpha-linolenic acid, linoleic acid, and conjugated linoleic
acid.
7. A pharmaceutical composition as in any one of claims 1-4 wherein
said polyunsaturated fatty acid is administered in the form of a
salt selected from the group consisting of a lithium salt, a sodium
salt, a potassium salt, a magnesium salt, a calcium salt, a
manganese salt, an iron salt, a copper salt, an aluminum salt, a
zinc salt, a chromium salt, a cobalt salt, a nickel salt and an
iodide.
8. A pharmaceutical composition as in any one of claims 1-4 wherein
said polyunsaturated fatty acid is in the form of a fatty acid
derivative selected from the group consisting of glycerides,
esters, free acids, amides, phospholipids and salts.
9. A pharmaceutical composition as in any one of claims 1-4 further
comprising an anti-neoplastic agent.
10. A pharmaceutical composition as in claim 9 wherein said
anti-neoplastic agent is selected from the group consisting of
tumor necrosis factor, an anti-cancer drug, a lymphokine, and
specific polyclonal or monoclonal antibodies.
11. A pharmaceutical composition as in claim 10 wherein said
lymphokine is selected from the group consisting of alpha
interferon and gamma interferon.
12. A pharmaceutical composition as in claim 10 wherein said
anti-cancer drug is selected from the group consisting of
vincristine, adriamycin, doxorubicin, cyclophosphamide,
cis-platinum, L-asparaginase, procarbazine, camptothecin, taxol and
busulfan.
Description
RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 09/946,129, filed Sep. 4, 2001, which is a
continuation-in-part of U.S. patent application Ser. No.
09/392,953, filed Sep. 9, 1999, the disclosures of which are
incorporated herein by reference.
BACKGROUND OF THE INVENTION
[0002] 1. Field of the Invention
[0003] This invention relates to methods for selectively occluding
blood vessels which supply neoplastic tissue, including tumors.
[0004] 2. Description of the Related Art
[0005] The polyunsaturated fatty acids (PUFAs) are fatty acids
having at least two carbon-to-carbon double bonds in a hydrophobic
hydrocarbon chain which typically includes X-Y carbon atoms and
terminates in a carboxylic acid group. The PUFAs are classified in
accordance with a short hand nomenclature which designates the
number of carbon atoms present (chain length), the number of double
bonds in the chain and the position of double bonds nearest to the
terminal methyl group. The notation "a:b" is used to denote the
chain length and number of double bonds, and the notation "n-x" is
used to describe the position of the double bond nearest to the
methyl group. There are 4 independent families of PUFAs, depending
on the parent fatty acid from which they are synthesized. They
are:
[0006] (1) The "n-3" series derived from alpha-linolenic acid (ALA,
18:3, n-3).
[0007] (2) The "n-6" series derived from linoleic acid (LA, 18:2,
n-6).
[0008] (3) The "n-9" series derived from oleic acid (OA, 18:1,
n-9).
[0009] (4) The "n-7" series derived from palmitoleic acid (PA,
16:1, n-7).
[0010] The parent fatty acids of the n-3 and n-6 series can not be
synthesized by the mammals, and hence they are often referred to as
"essential fatty acids" (EFAs). Because these compounds are
necessary for normal health but cannot be synthesized by the human
body, they must be obtained through the diet.
[0011] It is believed that both LA and ALA are metabolized by the
same set of enzymes. LA is converted to gamma-linolenic acid (GLA,
18:3, n-6) by the action of the enzyme delta-6-desaturase (d-6-d),
and GLA is elongated to form di-homo-GLA (DGLA, 20:3, n-6), the
precursor of the 1 series of prostaglandins. The reaction catalyzed
by d-6-d is the rate limiting step in the metabolism of EFAs. DGLA
can also be converted to arachidonic acid (AA, 20:4, n-6)) by the
action of the enzyme delta-5-desaturase (d-5-d). AA forms the
precursor of 2 series of prostaglandins, thromboxanes and the 4
series leukotrienes. ALA is converted to eicosapentaenoic acid
(EPA, 20:5, n-3) by d-6-d and d-5-d. EPA forms the precursor of the
3 series of prostaglandins and the 5 series of leukotrienes.
Conjugated linoleic acid (CLA; 18:2) is a group of isomers (mainly
9-cis, 11-trans and 10-trans, 12-cis) of linoleic acid. CLA is the
product of rumen fermentation and can be found in the milk and
muscle of ruminants (see, e.g., Brodie et al. (1999), J. Nutr. 129:
602-6; Visonneau et al. (1997), Anticancer Res. 17: 969-73. LA,
GLA, DGLA, AA, ALA, EPA, docosahexaenoic acid (DHA, 22:6, n-3) and
CLA are all PUFAs, but only LA and ALA are EFAs.
[0012] Under some well defined culture conditions GLA, AA, EPA and
DHA showed a marked differential cytotoxic effect against tumor
cells with little or no significant action on normal cells (Leary
et al. (1987), S. Afr. Med. J. 62:681-683; Begin et al. (1985),
Prostaglandins Leukot. Med. 19:177-186; Das (1999), Nutrition
15:239-241; Das (1991), Cancer Lett. 56:235-243; Das (1990),
Nutrition 6:429-434; Seigel et al. (1987), J. Natl. Cancer Inst.
78:271-277; Sangeetha and Das (1992), Cancer Lett. 63:189-198;
Begin et al. (1986), J. Natl. Cancer Inst. 77:1053-1062; Das
(1992), Asia Pacific J. Pharmacol. 7:305-327). In mixed culture
experiments, in which both normal and tumor cells were grown
together, GLA showed more selective tumoricidal action compared to
AA and EPA (Begin et al. (1986), Prog. Lipid Res. 25:573-576). In
addition, direct intra-tumoral administration of GLA can regress
human gliomas without significant side-effects (Naidu et al.
(1992), Prostaglandins Leukot. Essen. Fatty Acids 45: 181-184; Das
et al. (1995), Cancer Lett. 94:147-155).
[0013] Thus, it is known in the art that certain polyunsaturated
fatty acids (PUFAs) have cytotoxic properties towards tumor cells
in vitro, and that PUFAs provide the substrates for the generation
of lipid peroxidation products which have an inhibitory action on
cell proliferation. In addition, tumor cells are known to have low
d-6-d activity, which is necessary for the desaturation of LA and
ALA to their respective products. Moreover, it has been shown that
hepatocarcinogens, diethylnitrosamine (DEN) and
2-acetylamino-fluorine (2-AAF), can suppress the activity of d-6-d
and d-5-d resulting in low levels of GLA and AA, EPA and DHA in the
tumor cells.
SUMMARY OF THE INVENTION
[0014] In one aspect, the present invention provides methods of
selectively interrupting the blood supply to a neoplastic region,
such as a tumor region, causing necrosis of the neoplastic tissue
without substantial necrosis of adjoining tissues. The invention
also provides methods of selectively causing anti-angiogenic action
in a neoplastic region, such as a tumor region, with the result
that new blood vessels and collaterals are not formed to sustain
the neoplasia.
[0015] In particular, the invention provides methods for
selectively reducing blood supply to at least a portion of a
neoplastic region, in which (a) a proximal artery which carries
blood to at least a portion of said region is located and (b) a
therapeutically effective amount of a solution of at least one PUFA
is intra-arterially injected into the artery, thereby selectively
reducing the blood supply. In preferred embodiments, the amount of
the solution is sufficient to cause occlusion of the artery in a
period of less than one minute. In preferred embodiments, the
therapeutically effective amount is between 0.5 mg and 50 gm, most
preferably between 250 mg and 5 gm.
[0016] In some embodiments of the invention, in addition to the
PUFA, a lymphographic agent is intra-arterially injected to
visualize the proximal artery and blood supply to the neoplastic
region. The lymphographic agent may be combined with the PUFA
solution and they may be injected together. The progress of the
lymphographic agent through the proximal artery and neoplastic
region can be observed to determine when the blood supply is
effectively reduced and when injection of the PUFA solution can be
stopped. In some embodiments, the lymphographic agent is covalently
conjugated to the PUFA.
[0017] In some embodiments, the PUFA is an EFA. In certain
preferred embodiments, the EFA is selected from linoleic acid,
gamma-linolenic acid, arachidonic acid, docosahexaenoic acid,
eicosapentaenoic acid, di-homo-gamma-linolenic acid,
alpha-linolenic acid, linoleic acid, and conjugated linoleic
acid.
[0018] In preferred embodiments, the PUFA is administered in the
form of free acid or a salt, such as a lithium salt, a sodium salt,
magnesium salt, a manganese salt, an iron salt, a copper salt or an
iodide salt. In some preferred embodiments, the PUFA is in the form
of a fatty acid derivative, such as a glyceride, ester, ether,
amide, or phospholipid, or an alkylated, alkoxylated, halogenated,
sulfonated, or phosphorylated form of the fatty acid.
[0019] In some embodiments of the inventions, the neoplastic tissue
is a tumor. In particular, the neoplastic tissue may be a glioma,
hepatoma, lung cancer, colon cancer, breast cancer, ovarian cancer,
kidney cancer, skin cancer, Kaposi's sarcoma, esophageal cancer,
stomach cancer, leukemia, or lymphoma. In other embodiments, the
neoplastic tissue may result from a non-cancerous cell
proliferative disorder.
[0020] In some embodiments of the invention, in addition to the
PUFA, a therapeutically effective amount of a compound selected
from tumor necrosis factor, anticancer drugs, lymphokines, and
specific polyclonal or monoclonal antibodies is intra-arterially
injected. In preferred embodiments, the lymphokine is alpha
interferon or gamma interferon.
[0021] In some embodiments, the PUFA is covalently conjugated with
a pharmaceutical agent chosen from TNF, alpha-interferon,
gamma-interferon, an antibody, vincristine, adriamycin,
doxorubicin, cyclophosphamide, cis-platinum, L-asparaginase,
procarbazine, camptothecin, taxol or busulfan.
[0022] In another aspect, the invention provides pharmaceutical
compositions of a PUFA, or salt of PUFA, in combination with a
lymphographic agent or anti-neoplastic agent.
DETAILED DESCRIPTION
[0023] The patent, scientific and medical publications referred to
herein establish knowledge that was available to those of ordinary
skill in the art at the time the invention was made. The entire
disclosures of the issued U.S. patents, published and pending
patent applications, and other references cited herein are hereby
incorporated by reference.
[0024] Definitions.
[0025] In order to more clearly and concisely describe the subject
matter which is the invention, the following definitions are
provided for certain terms which are used in the specification and
appended claims.
[0026] As used herein, the term "neoplastic" means characterized by
abnormal tissue that shows partial or complete lack of structural
organization and functional coordination with normal tissue, and
usually forms a distinct mass which may be either benign or
malignant. As used herein, "neoplastic" tissue need not exhibit
cellular proliferation that is more rapid than normal tissue (e.g.,
a tumor which has ceased to grow or which is in remission).
Neoplastic tissue need not be cancerous (e.g., uterine fibroids,
adenomatous polyps of the intestines, adenomas in the lungs or
other organs).
[0027] As used herein, a "neoplastic region" means an essentially
contiguous region of tissue containing neoplastic tissue. A
neoplastic region is the smallest volume of tissue that includes
the contiguous neoplastic tissue, but may also include normal
tissue. Contiguous neoplastic tissues are neoplastic tissues
separated by distances of less than one centimeter, and do not
include distant metastases (which define separate neoplastic
regions). Although not all neoplastic regions are tumors, the terms
"neoplastic region" and "tumor" will often be used interchangeably
herein, and the term "tumorfeeding vessel" should be understood to
include an artery feeding any type of neoplastic region.
[0028] As used herein, the term "polyunsaturated fatty acid" and
the abbreviation "PUFA" mean any acid derived from fats by
hydrolysis, or any long-chain (at least 12 carbons) organic acid,
having at least two carbon-to-carbon double bonds. Examples of
PUFAs include but are not limited to linoleic acid, linolenic acid
and arachidonic acid.
[0029] As used herein, the term "PUFA salt" means an ionic
association, in solid or in solution, of a anionic form of a PUFA
with a cation of a small organic group (e.g., ammonium) or a small
inorganic group (e.g., an alkali metal). Preferred salts are those
between a PUFA and an alkali metal (e.g., lithium, sodium,
potassium), an alkali earth metal (e.g., magnesium, calcium) or a
multivalent transition metal (e.g., manganese, iron, copper,
aluminum, zinc, chromium, cobalt, nickel).
[0030] As used herein the term "lymphographic agent" means any of
the class of compounds which are used, or may be used, to visualize
lymphatics and lymph nodes, as well as veins and arteries,
following an intravenous or intra-arterial injection. Lymphographic
agents are typically vegetable oils (e.g., poppy seed oil) which
are iodized (e.g., approximately 30-45% by weight), and which may
be further derivatized (e.g., ethyl esterification). Examples
include the iodized fatty acids of poppy seed oil (commercially
available as LIPIODOL ULTRA FLUIDE.RTM. from Laboratoire Guerbet,
Paris, France), the ethiodized fatty acids of poppy seed oil
(commercially available as ETHIODOL.RTM. from Savage Laboratories,
Melville, N.Y.) and iophendylate (PANTOPAQUE.RTM. from Kodak). See,
Hom et al. (1957), J. Am. Pharm. Assoc. Sci. Ed. 46:254; Paxton et
al. (1975), Brit. Med. J. 1:120. As used herein, the term
"lymphographic agent" means any agent which is useful for
non-invasively visualizing blood vessels including, without
limitation, radiography, CAT scans, MRI scans, ultrasound imaging,
and the like.
[0031] As used herein, the term "angiogram" means any method of
noninvasively visualizing a blood vessel or lymphatic including,
without limitation, radiography, CAT scans, MRI scans, ultrasound
imaging, and the like.
[0032] As used herein, the term "proximal" is a relative term which
describes the location of an artery with respect to a neoplastic
region and a site of intra-arterial injection of a PUFA salt of the
invention. An artery is proximal to a neoplastic region if it is
upstream of the neoplastic region with respect to blood flow and
downstream (or distal) of the site of injection with respect to
blood flow. A proximal artery should also be physically close to
the neoplastic region such that a substantial portion (e.g., at
least 10%, preferably 25%, most preferably greater than 50%) of the
volume of a solution injected into the artery would normally pass
into arteries, arterioles and capillary beds within the neoplastic
region.
[0033] General Considerations
[0034] The present invention is dependent, in part, upon the
discovery of the novel and highly beneficial action of PUFAs, and
especially certain PUFA salts, to induce the selective occlusion of
blood vessels feeding neoplastic regions, including tumors. This
effect is particularly observed when the PUFA is administered in
combination with a lymphographic agent comprising iodized fatty
acids.
[0035] Without being bound to any particular theory of the
invention, it is believed that the selective occlusion of the
tumor-feeding vessels is not due to embolism or other forms of
physical blockage. This conclusion follows from observations in
several patients that normal blood vessels, which were sometimes
smaller in diameter than tumor-feeding vessels, which were located
proximal to the tumor-feeding vessels, and which were closer to the
tip of the catheter and the site of injection, were not occluded.
If the occlusion were due to embolization, all blood vessels,
especially those that were smaller in diameter compared to the
tumor-feeding vessels, would be expected to be occluded first.
Because the site of injection of the PUFA, as determined by
angiographic imaging of the tip of the catheter in several
patients, was slightly upstream from the origin of the main
tumor-feeding vessels, it is evident that the occlusion of the
tumor-feeding vessels is not due to direct injection of the PUFA
only into those vessels. Rather, the ability of PUFA to selectively
occlude the tumor-feeding vessels but not normal arterial vessels
was seen in several patients.
[0036] Moreover, in several other patients, a PUFA was injected
into normal arteries including the celiac, subclavian and popliteal
arteries. During the course of these procedures, no spasms or
occlusions (even temporary) of these blood vessels were observed.
On the other hand, the PUFA occluded all types of tumor-feeding
vessels, irrespective of their size, almost instantaneously.
Without being bound to any particular theory of the invention, this
rapid action of the PUFA suggests that an intense vasospasm was
induced (directly or indirectly) in the tumor-feeding vessels but
not in the normal blood vessels and that, following such a
vasospasm, thrombosis may have led to permanent occlusion of the
blood vessel.
[0037] Finally, without being bound to any particular theory of the
invention, it is believed that there is an interaction between the
PUFA and lymphographic agents of the invention which may account
for the effectiveness of the treatment. Thus, lymphographic agents
comprising iodized fatty acids, and particularly the iodized fatty
acids of vegetable oils, are believed to synergistically interact
with the PUFAs to produce a therapeutic effect which is
qualitatively different than the effect of either the PUFA or the
lymphographic agent alone.
[0038] There are several advantages of PUFA treatments of the
invention. As shown below, a single injection (or at most two or
three injections at separate times, if the neoplastic region is
large) is adequate to produce almost permanent occlusion of the
tumor-feeding vessels. The PUFAs and their salts are non-antigenic,
are known to be relatively safe in the dosages employed, and are
stable. The dosage of PUFA needed to occlude the tumor-feeding
vessels in a given patient is self-evident during administration:
As the PUFA solution is being injected, and as the tumor-feeding
vessels are being occluded, resistance to further injection will be
felt, at which point the injection can be stopped.
[0039] The invention in one aspect provides methods of inhibiting
blood supply to a neoplastic region, comprising the steps of (a)
locating an artery which carries major blood supply to the
neoplastic region and which is proximal to the neoplastic region;
and (b) intra-arterially injecting into the located artery a
solution of at least one PUFA chosen from LA, GLA, DGLA, AA, ALA,
EPA, DHA and CLA. In preferred embodiments, the PUFA is
administered in combination with a lymphographic agent.
[0040] The invention in another aspect provides methods for
treating neoplasias and for facilitating the visualization of
remission of a neoplasia which is responsive to treatment,
comprising the steps of (a) locating an artery proximal to the
neoplastic region which carries a major portion of blood supply to
the neoplastic region and which is adjacent to the neoplastic
region; (b) obtaining an initial radiographic image of the region;
(c) injecting into the artery a mixture of (i) a lymphographic
agent, and (ii) a solution of at least one PUFA chosen from LA,
GLA, DGLA, AA, ALA, EPA, DHA and CLA; and (d) obtaining second and,
optionally, subsequent radiographic images of the neoplastic region
after predetermined lapses of time; and comparing the initial
radiographic images with the second and/or subsequent radiographic
images to assess the extent of remission of the neoplasia.
[0041] The invention in another aspect provides methods of causing
necrosis in a neoplastic region (e.g., a cancerous tumor) by
inhibiting blood supply to the neoplastic region, comprising the
steps of (a) locating an artery proximal to the neoplastic region
which carries major blood supply to the neoplastic region; (b)
injecting into the located artery a mixture of (i) a lymphographic
agent, and (ii) a solution of at least one PUFA chosen from LA,GLA,
DGLA, AA, ALA, EPA, DHA and CLA; (c) waiting for a predetermined
time period and assessing a degree of necrosis in the neoplastic
region; and (d) repeating the treatment if necessary to increase
the necrosis.
[0042] In yet another aspect, the invention provides methods of
treating mammalian cell proliferative disorders using a solution of
a PUFA, or combinations of PUFAs, administered intra-arterially.
The methods are as described above with respect to neoplastic
regions.
[0043] In each of the foregoing embodiments, the PUFA is preferably
in the form of a salt, most preferably in the form of a lithium
salt, and is preferably administered in combination with a
lymphographic agent. The lymphographic agent is preferably an
iodized fatty acid derived from a vegetable oil.
[0044] Although the invention is described primarily as it relates
to humans, it is envisaged that the methods of the invention are
equally applicable to other mammals, including large domesticated
mammals (e.g., race horses, breeding cattle) and smaller
domesticated animals (e.g., house pets).
[0045] Choice of PUFA
[0046] The present invention employs PUFAs, preferably in the form
of salts, to selectively occlude arteries which provide blood
supply to regions of neoplastic tissue. Preferred PUFAs include,
but are not limited to, GLA, AA, DHA, EPA, DGLA, ALA, LA and CLA.
Other preferred PUFAs include derivatives of the aforementioned
PUFAs, including glycerides, esters, ethers, amides, or
phospholipids, or alkylated, alkoxylated, halogenated, sulfonated,
or phosphorylated forms of the fatty acid. In most preferred
embodiments, the PUFA is GLA, AA or DHA.
[0047] The PUFA is preferably administered in the form of a salt
solution. Suitable salts include salts of a PUFA with a cation of a
small organic group (e.g., ammonium) or a small inorganic group
(e.g., an alkali metal or alkali earth metal). Preferred referred
salts are those between a PUFA and an alkali metal (e.g., lithium,
sodium, potassium), an alkali earth metal (e.g., magnesium,
calcium) or a multivalent metal (e.g., manganese, iron, copper,
aluminum, zinc, chromium, cobalt, nickel). Most preferred are salts
of lithium, sodium, magnesium, manganese, iron, copper, and
iodides. Combinations of salts may also be employed.
[0048] When the PUFAs or PUFA salts are administered in combination
with an oily lymphographic agent or other agents, the solution may
be formed into an emulsion.
[0049] Lymphographic Agents
[0050] In order to visualize lymphatic vessels, lymph nodes,
arteries and veins, lymphographic agents are frequently employed.
In the context of the present invention, these agents may aid in
both the placement of a syringe or catheter in a proximal artery
for intra-arterial injection of a PUFA solution, and may also aid
in the visualization of the resulting selective occlusion of
tumor-feeding vessels. In addition, such agents may be used in
follow-up angiograms to determine whether occlusion has been
successful or complete, and to determine whether additional
treatments may be necessary. Finally, it is believed that such
agents have a synergistic or potentiating effect in combination
with the PUFA solutions of the invention, and thereby serve as
additional or ancillary active ingredients in the treatments of the
invention.
[0051] The lymphographic agents can be any of the class of
compounds, recognized by those of skill in the art of diagnostic
imaging, which are used, or which may be used, to visualize
lymphatics and lymph nodes, as well as veins and arteries, by
radiography following an intra-lumenal injection. Lymphographic
agents are typically vegetable oils (e.g., poppy seed oil) which
are iodized (e.g., approximately 30-45% by weight), and which may
be further derivatized (e.g., ethyl esterification). Examples
include the iodized fatty acids of poppy seed oil (commercially
available as LIPIODOL ULTRA FLUIDE.RTM. from Laboratoire Guerbet,
Paris, France), the ethiodized fatty acids of poppy seed oil
(commercially available as ETHIODOL.RTM. from Savage Laboratories,
Melville, N.Y.) and iophendylate (PANTOPAQUE.RTM. from Kodak). See,
Hom et al. (1957), J. Am. Pharm. Assoc. Sci. Ed. 46:254; Paxton et
al. (1975), Brit. Med. J. 1:120.
[0052] The lymphographic agents of the invention may be mixed with
the PUFA solutions described above, either to form a new solution
or to form an emulsion, or they may be chemically conjugated to the
PUFAs of the invention via standard chemistries. Preferably the
lymphographic agent is an iodized lymphographic oil, such as an
iodized poppy seed oil. Preferably the PUFA solution is mixed with
such a lymphographic agent in a ratio of at least about 2:1, or
about 1:1, or about 1:1.5, or about 1:2, or about 1:3
(volume/volume). Most preferably the ratio is between 1:1.5 and 1:3
(volume/volume). The preferred lymphographic agent is LIPIODOL
ULTRA FLUIDE.RTM. (Laboratoire Guerbet, Paris, France). This
lymphographic agent may be safely administered to a typical patient
in an amount of about 10 mL/m.sup.2, but the attending physician
should consider all relevant medical factors in determining the
appropriate dosage for any specific patient.
[0053] Thus, in another aspect, the invention provides
pharmaceutical compositions comprising a PUFA, or a PUFA salt, and
a lymphographic agent in solution, or in an emulsion. The PUFA and
lymphographic agent may be separate chemical moieties combined in
the solution or emulsion, or they may be covalently conjugated. The
preferred lymphographic agents and ratios for such a product are as
disclosed above. Preferably the final concentration of the PUFA in
such a product is at least 5%, preferably at least 20%, and most
preferably about 25-50%.
[0054] Methods of Administration
[0055] The PUFA solutions of the present invention are preferably
administered intra-arterially to an artery which is proximal to the
neoplastic region to be treated. The approximate location of the
neoplastic region must first be identified by any of the methods
known in the art. For example, X-rays, Computerized Axial
Tomography (CAT) scans, Magnetic Resonance Imaging (MRI) scans,
palpation or direct visual inspection may be used to identify a
neoplastic region. Such methods may optionally employ contrast
agents, including lymphographic agents or agents specifically
targeted to neoplastic tissues (e.g., radioisotope-labeled
antibodies against tumor-associated antigens). Once the neoplastic
region is identified, an artery which feeds the region (i.e., which
is upstream with respect to blood flow to the region) is
identified. The intra-arterial injection site is preferably chosen
to be close or proximal to the neoplastic region to increase the
portion of the dosage which reaches that region, but is also
preferably chosen sufficiently far upstream from that region such
that all or most of the neoplastic region receives a portion of the
injected dosage.
[0056] Thus, as one progresses along an artery which feeds a
neoplastic region, the artery will branch into smaller and smaller
arteries and finally arterioles. At some distant point upstream
from the neoplastic region, the artery will feed not only the
neoplastic region but also large regions of normal tissue. As the
chosen injection site is moved along the artery toward the
neoplastic region, the percentage of blood carried by the artery
which feeds normal tissue will decrease. By proceeding along the
artery toward the neoplastic region, therefore, one can increase
the portion of the dosage which reaches the neoplastic region.
However, as the injection site proceeds along the artery toward the
neoplastic region, one may also bypass branches of the artery which
feed the neoplastic region and, therefore, fail to cause occlusion
of arteries supplying a part of the neoplastic region. One of
ordinary skill in the art may balance these considerations, as well
as other considerations (e.g., accessibility of an artery for
catheterization), in choosing a site for injection. Thus, the term
"proximal" is a relative term which describes the location of an
artery with respect to a neoplastic region and a site of
intra-arterial injection of a PUFA of the invention. Preferably, a
proximal artery should also be physically close to the neoplastic
region such that a substantial portion (e.g., at least 10%,
preferably 25%, and most preferably greater than 50%) of the volume
of a solution injected into the artery would normally pass into
arteries, arterioles and capillary beds within the neoplastic
region. Thus, the hepatic artery might be considered proximal to a
neoplastic region in the liver, but the descending aorta would
not.
[0057] In order to administer a PUFA solution to a proximal artery,
the artery is identified as described above, the site of injection
is chosen, and the PUFA solution is administered by injection
through a syringe or catheter as appropriate to the location. As
necessary, the syringe or catheter may be guided to the site of
injection by radiological guidance (e.g., X-rays), CAT guidance,
MRI guidance, endoscopic guidance, or stereotaxic guidance. In the
case of a catheter, the catheter can be inserted into the body at a
site quite distant from the proximal artery, and then be guided to
the proximal artery. For example, the femoral, brachial and carotid
arteries may provide convenient entry points for a catheter which
is then routed to a proximal artery elsewhere in the body. In
addition, contrast agents may be added to the injected solution to
aid in placement of the syringe or catheter, or to aid in
visualization of the occlusion of the tumor-feeding vessels.
[0058] Appropriate dosages of the PUFA solutions of the invention
will depend primarily on the diameter of the proximal artery at the
site of injection and the number and size of the arteries and/or
arterioles branching therefrom. Preferred dosages range from
approximately 0.5 mg for the smallest proximal arteries to 50 gm
for very large proximal arteries feeding large neoplastic regions.
More typically, dosages of approximately 250 mg to 5 gm are
preferred and, as shown in the examples below, dosages of 500 mg to
750 mg are effective for several different types of tumors.
However, in most preferred embodiments of the methods of the
invention, the PUFA solution is administered in combination with a
lymphographic agent, the administration is observed by angiogram,
and administration continues until the tumor-feeding vessels are at
least partially occluded as indicated by the angiogram.
Alternatively or additionally, administration may be continued
until a significant increase in resistance to the injection
develops, indicating the tumor-feeding vessels distal to the site
of injection have been at least partially occluded.
[0059] Other Agents
[0060] The PUFA solutions of the invention may be administered
alone, or in combination with other pharmaceutical agents known in
the art for the treatment of neoplasias. Thus, for example, the
PUFA solutions may be co-administered with known anti-cancer drugs,
including vincristine, adriamycin, doxorubicin, cyclophosphamide,
cisplatinum, L-asparaginase, procarbazine, camptothecin, taxol and
busulfan. Alternatively, the PUFA solutions may be co-administered
with known lymphokines such as tumor necrosis factor (TNF) and/or
an interferon (e.g., alpha interferon or gamma interferon) or
specific polyclonal or monoclonal antibodies.
[0061] Administration of these agents in combination with a PUFA
solution, or a PUFA and lymphographic agent solution, may also show
a synergistic or potentiating effect.
[0062] Thus, in another aspect, the invention provides
pharmaceutical compositions comprising a PUFA, or a PUFA salt, and
a pharmaceutical agent known in the art for the treatment of
neoplasias, either in solution, or in an emulsion. The PUFA and
other pharmaceutical agent may be separate chemical moieties
combined in the solution or emulsion, or they may be covalently
conjugated. The preferred pharmaceutical agents are as disclosed
above. Preferably the final concentration of the PUFA in such a
product is at least 5%, preferably at least 15%, and most
preferably at least 25%. The product may contain substantially more
PUFA, up to 100% without any significant side-effects.
[0063] The following examples illustrate some preferred modes of
practicing the present invention, but are not intended to limit the
scope of the claimed invention. Alternative materials and methods
may be utilized to obtain similar results.
EXAMPLES
[0064] Patients
[0065] Studies were conducted in 5 human patients with stage 4
neoplastic disease. Two of the patients had primary hepatoma
(patients 1 and 2), two had giant cell tumor of the bone (patients
3 and 4) and one had renal cell carcinoma (patient 5).
[0066] Materials
[0067] The lithium salt of GLA was obtained from Scotia
Pharmaceuticals, U.K. The PUFA salt was dissolved in sterile
saline, sterile phosphate buffered saline (PBS, pH 7.4) or dilute
ethanol in saline (final concentration <0.02% ethanol). The
final concentration of PUFA in these solutions was approximately
25%. The PUFA solution was mixed with an iodized lymphographic oil
(LIPIODOL ULTRA FLUIDE.RTM., Laboratoire Guerbet, Paris, France),
in a ratio of between 1:1.5 and 1:3 (volume/volume). In some cases,
the PUFA was modified by covalent conjugation (e.g., amide bond) to
the iodized lymphographic oil. In other cases the PUFA salt was
unmodified, and was diluted directly into the lymphographic agent
without conjugation. The PUFA and lymphographic oil mixture was
prepared under sterile conditions immediately prior to use.
[0068] Methods of Administration
[0069] Patients were admitted into the hospital for the study. A
proximal artery supplying a major portion of the blood supply to
the tumor was identified. Catheterization of the major artery from
which the proximal artery arises was performed under local
anaesthesia. In patients 1 and 2 (with hepatomas), the tip of the
catheter was positioned in the right hepatic artery via the right
femoral artery. In patient 3 (with giant cell tumor of the right
lower end of the femur), the tip of the catheter was positioned in
the right femoral/popliteal artery. In patient 4 (with giant cell
tumor of the left scapula), the tip of the catheter was positioned
in the left subclavian/axillary arteries. In patient 5 (with left
kidney tumor), the tip of the catheter was positioned in the left
renal artery. Conjugated PUFA salt was prepared fresh, just prior
to injection. Radiographic and CT scan examinations were performed
immediately after the injection and at periodic intervals in all
the five patients.
[0070] In order to determine how the arterial supply to the tumor
tissue was influenced by the injection of PUFA salt, angiography
was performed and recorded during and immediately after the
procedure, and at periodic intervals thereafter.
[0071] Patients 1 and 2 with hepatoma were administered total doses
of 1.6 gm and 0.75 gm respectively (the dose refers to the amount
of Li-GLA) of PUFA salt into the right hepatic artery through the
right femoral route. Patient 3, with giant cell tumor of the lower
end of the right femur, underwent right femoral artery
catheterization and the tip of the catheter was positioned in the
popliteal artery to deliver 500 mg of PUFA salt. Patient 4, who had
giant cell tumor of the left scapula, received PUFA salt by
selective cannulation of the left subclavian artery, through the
femoral route, from which the tumor-feeding vessel was arising and
was given 660 mg of PUFA salt. Patient 5, with renal tumor,
received 750 mg of PUFA salt through the right femoral route into
the left renal artery from which the tumor-feeding vessels were
arising. In all the patients, the administration of PUFA salt was
done as swiftly as possible. During the administration of PUFA
salt, the vital signs of each patient were monitored.
[0072] Summary Results
[0073] All 5 patients tolerated the treatment well and no
significant side-effects due to the therapy were noted. The only
side-effect was a complaint of a mild feeling of warmth followed by
pain at the site of the tumor during and immediately after the
injection of the PUFA salt. This was presumably due to the
perfusion of the tumor with the PUFA salt, and ischaemia resulting
from occlusion of the tumor-feeding vessels. In general, the pain
was not severe and was ameliorated by the administration of
non-steroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen
or buscopan. All the biochemical tests performed after the
administration of the PUFA salt were found to be normal.
[0074] The most significant and surprising observation of these
studies was the total occlusion of the tumor-feeding vessels
following PUFA salt injection. This was a consistent observation in
all the 5 patients. This selective occlusion of the tumor-feeding
vessels was seen during the course of injecting the PUFA salt in
the patients with giant cell tumors of the bone and the patient
with renal cell carcinoma. In the case of the patients with
hepatomas, the occlusion of the tumor-feeding vessels was noticed
over a period of time. In patient 1 the occlusion of the
tumor-feeding vessels was observed 10 days after the first
injection of PUFA salt, and six days after a second injection, but
presumably occurred during or shortly after the second injection.
No occlusion was seen in the normal vasculature feeding
non-neoplastic tissues. The time lag for the occlusion of the
tumor-feeding vessels observed in patient 1 with hepatoma suggests
that the occlusion of the tumor-feeding vessels is not due to an
embolization process. Moreover, normal blood vessels, which were
much smaller in diameter compared to the tumor-feeding vessels,
were not occluded when exposed to PUFA salt, further suggesting
that embolization was not the mechanism. On the other hand, in
patient 2, also with hepatoma, the occlusion of the tumor-feeding
vessels was seen essentially simultaneously with the injection of
the PUFA salt. The remarkably selective occlusion of only the
tumor-feeding vessels was clear from pre- and post-injection
angiograms of patient 5 with the renal cancer. These angiograms
showed that the three main tumor-feeding vessels arising from the
main renal artery were completely occluded whereas the fourth and
the fifth branches arising from the main stem of the left renal
artery, which were feeding the normal lower pole of the kidney,
were not occluded. This was despite the fact that all the five
vessels were arising from the same renal artery, downstream from
the point of injection. If the occlusion of the vessels were the
result of embolization, it is expected that the lower most and
narrower branches should have been occluded first and later the
other vessels.
[0075] The pre and post-injection angiograms of all the 5 patients
indicated that PUFA salt can selectively occlude tumor-feeding
vessels. In order to know the duration of this selective occlusion
of the tumor-feeding vessels, angiograms were repeated at periodic
intervals. It was noted that even 28 days after PUFA salt
injection, no tumor-feeding vessels could be seen in patient 1 with
hepatoma. In the patient with giant cell tumor of the lower end of
the right femur (patient 3), a repeat angiogram performed after 10
days following PUFA salt injection also did not show any
tumor-feeding vessels. Radiographs of the right knee of this
patient taken 10 days after the injection clearly showed the
presence of the contrast material, which is due to the presence of
conjugated PUFA salt in the tumor tissue. In the patient with giant
cell tumor of the left scapula (patient 4), a follow up angiogram
performed seven and a half years after the injection of PUFA salt
clearly showed that the original tumor-feeding vessel was still
occluded. A plain radiograph of the left scapula showed extensive
sclerosis of the tumor, and attempts to do a fine needle aspiration
biopsy of the healed mass were unsuccessful due to its hard, bony
nature.
[0076] Detailed Results
[0077] Patient 1. Patient 1 was a 45 year old male. The patient was
diagnosed to have hepatoma of the right lobe of the liver confirmed
by fine needle aspiration biopsy (FNAB). The patient was considered
unsuitable for surgery, radiotherapy and chemotherapy due to poor
general condition and large size of the tumor. The patient had lost
more than 25 kgs prior to confirmation of the diagnosis. Prior to
the treatment, a pronounced "tumor blush" was observed in
angiograms. In a first treatment, 1.1 gm of PUFA salt was
administered. More than 50% of the tumor-feeding vessels were
occluded following the first injection, and a significant amount of
resistance was noted while injecting the first dose of PUFA salt. A
repeat angiogram done 4 days after the first dose of the PUFA salt
showed that the tumor blush was much reduced. At that time, an
additional dose of 0.5 gm of PUFA was administered. A third
angiogram performed 1 week after the second dose of PUFA salt (and
the 11th day after the first dose) showed almost complete occlusion
of the tumor-feeding vessels. No such occlusion was seen in the
normal vasculature feeding non-neoplastic tissues. A >50%
reduction in the size of the hepatoma in patient 1 was seen one
month after the first PUFA salt injection. The decrease in the size
of hepatoma was associated with an increase in the radiographic
density of the contrast agent. This suggests that, as the size of
the tumor was decreasing, there was a concomitant increase in the
density of the contrast agent (to which the PUFA salt was
conjugated) in the remaining portion of the tumor. The patient felt
well for 6 months following the injection, gained >10 kgs in
weight, and was asymptomatic. Unfortunately, the patient died in a
traffic accident.
[0078] Patient 2. Patient 2 was a 50 year old male. The patient was
diagnosed to have hepatoma of the right lobe of the liver confirmed
by FNAB, and was considered to be high risk for surgery, and
unlikely to respond to radiotherapy and/or chemotherapy. During the
course of injecting the PUFA salt, significant resistance was felt
and therefore only 750 mg of PUFA salt was administered into the
right hepatic artery. An angiogram recorded immediately after the
injection showed complete occlusion of the tumor-feeding vessels.
The patient experienced a mild to moderate degree of pain in the
hepatic region during and after the injection, which subsided after
administering NSAIDs (ibuprofen). The pain subsided after 24 to 48
hours. This patient developed peritonitis 1 week after the PUFA
administration. Subsequent investigations revealed that the patient
had developed a perforated duodenal ulcer, which was responsible
for the peritonitis but presumably unrelated to the PUFA injection.
The patient died on the 10th day due to complications of the
perforated duodenal ulcer.
[0079] Patient 3. Patient 3 was a 24 year old male. The patient was
diagnosed to have giant cell tumor (osteoclastoma) involving the
medial condyle of the right femur. The diagnosis was confirmed by
biopsy. The patient refused the recommended surgical amputation
above the right knee, and opted to try a PUFA injection. A total
dose of 500 mg of PUFA salt was delivered into the
femoral/popliteal artery. Complete occlusion of the tumor-feeding
vessels was noted immediately after the injection. The patient
complained of pain in the right knee area and was given NSAIDs. A
repeat angiogram done 10 days after the injection of PUFA salt
showed that the occluded tumor-feeding vessels remained closed. The
patient appeared for a follow-up 2 months after the injection, at
which time he reported that he was better except for mild pain in
the region of the tumor. The patient was subsequently lost for
follow up visits.
[0080] Patient 4. Patient 4 was a 30 year old male. The patient was
diagnosed to have giant cell tumor of the left scapula, confirmed
by biopsy. The patient refused the recommended surgical removal of
the left upper limb and opted to try a PUFA injection. The patient
received 660 mg of PUFA salt through selective catheterization of
the left subclavian artery. During the course of the injection,
significant resistance was felt and the injection was stopped after
delivering 660 mg of PUFA salt. Complete occlusion of the
tumor-feeding vessels was seen immediately after the administration
of PUFA salt. This patient was discharged from the hospital after
24 hours of observation. The patient felt better and was able to do
use his left arm normally, without any pain or limitation of
movement. Seven and a half years after the PUFA salt injection, an
angiogram showed that the tumor-feeding vessels were still occluded
and that the tumor mass was completely calcified. Attempts at FNAB
were not successful due to hard nature of the healed mass.
[0081] Patient 5. Patient 5 was a 79 year old male. This patient
was diagnosed to have right renal cell carcinoma. By the time it
was diagnosed, the disease was well advanced, with secondary tumors
in the liver, peritoneum and right pleural effusion. The patient
had lost about 10 kgs of weight and was considered a high risk for
surgery. In a first treatment, 750 mg of PUFA salt was delivered
into the right renal artery via the right femoral route. Complete
occlusion of the tumor-feeding arteries was noted without any
occlusion of the normal arteries which were feeding the normal
lower pole of the right kidney. The patient experienced mild to
moderate pain at the site of tumor (right lumbar area). The pain
subsided after administering NSAIDs. The patient complained of pain
in the right knee area and was given NSAIDs (buscopan by injection
and oral ibuprofen). The patient was sent home 48 hours after PUFA
salt injection. The patient survived for 3 months, and then died
due to extensive metastasis in the liver and lungs.
[0082] Equivalents
[0083] While this invention has been particularly shown and
described with references to preferred embodiments thereof, it will
be understood by those skilled in the art that various changes in
form and details may be made therein without departing from the
spirit and scope of the invention as defined by the appended
claims. Those skilled in the art will recognize, or be able to
ascertain using no more than routine experimentation, many
equivalents to the specific embodiments of the invention described
specifically herein. Such equivalents are intended to be
encompassed in the scope of the appended claims.
* * * * *