U.S. patent application number 15/635507 was filed with the patent office on 2017-12-28 for par-4 agonists for the treatment of cancer.
The applicant listed for this patent is Cipla Limited. Invention is credited to Jeevan Ghosalkar, Kalpana Joshi, Geena Malhotra.
Application Number | 20170368072 15/635507 |
Document ID | / |
Family ID | 60675176 |
Filed Date | 2017-12-28 |
United States Patent
Application |
20170368072 |
Kind Code |
A1 |
Malhotra; Geena ; et
al. |
December 28, 2017 |
PAR-4 AGONISTS FOR THE TREATMENT OF CANCER
Abstract
Disclosed herein are methods of treating cancers by
administration of a PAR-4 agonist.
Inventors: |
Malhotra; Geena; (Dadar
(East), IN) ; Joshi; Kalpana; (Maharashtra, IN)
; Ghosalkar; Jeevan; (Thane, IN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Cipla Limited |
Mumbai |
|
IN |
|
|
Family ID: |
60675176 |
Appl. No.: |
15/635507 |
Filed: |
June 28, 2017 |
Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61K 45/06 20130101;
A61K 31/546 20130101 |
International
Class: |
A61K 31/546 20060101
A61K031/546; A61K 45/06 20060101 A61K045/06 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 28, 2016 |
IN |
201621022084 |
Claims
1. A method for treating cancer, comprising administering
cefuroxime, a pharmaceutically acceptable salt or prodrug thereof,
to a patient in need thereof, in an amount effective to increase
expression of prostate apoptosis response 4 (PAR-4).
2. The method according to claim 1, wherein the cefuroxime prodrug
comprises cefuroxime axetil.
3. The method according to claim 1, wherein the method further
comprises administering at least one additional cancer
treatment.
4. The method according to claim 3, wherein the additional cancer
treatment comprises administering one or more additional
chemotherapeutic agents.
5. The method according to claim 3, wherein the additional cancer
treatment comprises administering ionizing radiation to the
patient.
6. The method according to claim 5, wherein the cefuroxime is
administered for a period of at least one week prior to commencing
radiation treatment.
7. The method according to claim 3, wherein the additional cancer
treatment comprises a surgical intervention.
8. The method according to claim 7, wherein the cefuroxime is
administered for a period of at least one week prior to the
surgical intervention.
9. The method according to claim 4, wherein the cefuroxime and
additional chemotherapeutic acid are administered in an
intermittent dosing regimen.
10. The method according to claim 4, wherein the cefuroxime and
additional chemotherapeutic acid are administered
simultaneously.
11. The method according to claim 4, wherein the cefuroxime and
additional chemotherapeutic acid are administered separately.
12. A pharmaceutical composition comprising cefuroxime and at least
one additional therapeutic agent.
13. The composition according to claim 12, wherein the additional
therapeutic agent is a chemotherapeutic agent.
14. A kit comprising cefuroxime and at least one other therapeutic
agent.
15. The kit according to claim 17, wherein the additional
therapeutic agent is a chemotherapeutic agent.
Description
CROSS REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of Indian Application
201621022084, filed on Jun. 28, 2016, the contents of which are
hereby incorporated in its entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to the treatment of cancer,
specifically to a method of treating cancer by administering a
PAR-4 agonist to a patient in need thereof.
BACKGROUND
[0003] Cancer is a major public health problem in the United States
and many other parts of the world. It is currently the second
leading cause of death in the United States, and is expected to
surpass heart diseases as the leading cause of death in the next
few years. It remains a major cause of mortality in the world.
Despite the improvements that have been made in therapies and in
understanding the molecular basis of cancer, mortality is still
high. The current treatment regimens for cancer have shown limited
survival benefits when used for most advanced stage cancers.
[0004] The research and efforts being invested for cancer treatment
has changed over the past few decades. The age when surgery and
radiotherapy were the only effective way to fight tumor growth has
ended. A complex scenario where the molecular features of tumors
seem to be the cornerstone of any therapy is now emerging with new
targets or receptors being discovered in vivo. Continued research
has expanded knowledge of how cancer develops and how to target
medicines for specific cancer types, which has resulted in more
effective therapies for patients. However these therapies show a
lack of efficacy in terms of long-term outcome because of their
failure to target cancer cells and lead to toxicity due to
non-specific effects on normal cells. To overcome these side
effects, researchers have tried to understand the root cause and
have explored more about the gene changes in cells that cause
cancer, they have been able to develop drugs that target these
changes. Targeted therapy drugs does not work in the same way as
the standard chemotherapy drugs. They are often able to attack
cancer cells while doing less damage to normal cells by targeting
the programming of cancer cells that sets them apart from normal,
healthy cells. These drugs tend to have different (and often less
severe) side effects than standard chemotherapy drugs. Examples of
the targeted therapies include sorafenib, sunitinib, bevacizumab,
telomerase etc.
[0005] Research on apoptosis has increased substantially since the
early 1990s. Apoptosis (programmed cell death)-inducing drugs
change proteins within the cancer cells and cause the cells to
die.
[0006] Apoptosis can be initiated through one of two pathways. In
the intrinsic pathway the cell kills itself because it senses cell
stress, while in the extrinsic pathway the cell kills itself
because of signals from other cells. Both pathways induce cell
death by activating caspases, which are proteases, or enzymes that
degrade proteins. The two pathways both activate initiator
caspases, which then activate executioner caspases, which then
cause cell apoptosis by degrading proteins indiscriminately.
[0007] Induction of apoptosis in malignant cells therefore becomes
a major goal of cancer therapy in general and of certain cancer
gene therapy strategies in particular. Numerous
apoptosis-regulating genes have been evaluated for this purpose for
example p53 gene, p16, p21, p27, E2F genes, FHIT, PTEN, E1A and
CASPASE genes.
[0008] The prostate apoptosis response-4 (PAR-4) gene was first
identified by the differential hybridization technique as an
immediate early apoptotic gene upregulated in response to elevated
intracellular Ca2+ concentration [Ca2+] in the androgen-independent
rat prostate cancer cells AT-3 treated with ionomycin (Sells SF, et
al; Cell Growth Differ. 1994 Apr.; 5(4):457-66).
[0009] Studies conducted in cell culture models show that
over-expression of PAR-4 is sufficient to directly induce apoptosis
in many cancer cell types. The ability of PAR-4 to directly cause
apoptosis is associated with its nuclear translocation. Moreover,
the apoptotic action of PAR-4 can overcome cell protective
mechanisms, such as the presence of Bcl-xL, Bcl-2, or absence of
wild-type p53 or PTEN function. Interestingly, PAR-4 is incapable
of directly inducing apoptosis in normal or immortalized normal
cells.
[0010] The "apoptosis-sensitizing" function of PAR-4 in some of the
cancer cells is attributed to its accumulation in the cytoplasm and
inability to translocate into the nucleus, due to phosphorylation
by Aktl which renders PAR-4 subject to sequestration in the
cytoplasm by complexing it with chaperone proteins such as 14-3-3
(Goswami et al., Mol Cell. 2005 Oct. 7; 200:33-44.); however,
treatment with the other apoptotic signals translocates PAR-4 into
the nucleus to produce apoptosis.
[0011] Thus there is a need for improved methods for treating
cancer. There remains a need for selective PAR-4 agonists. There
remains a need for identifying selective PAR-4 agonists useful in
the treatment of cancer.
SUMMARY
[0012] According to an aspect of the present invention, there is
provided a method for enhancing or promoting PAR-4 expression by
administering a PAR-4 agonist. In some instances the PAR-4 agonist
is a cephalosporin antibiotic, for instance a second generation
cephalosporin antibiotic such as cefuroxime, salts and prodrugs
thereof.
[0013] According to an aspect of the present invention, there is
provided a method of treating cancer comprising administering by
administering a PAR-4 agonist. In some instances the PAR-4 agonist
is a cephalosporin antibiotic, for instance a second generation
cephalosporin antibiotic such as cefuroxime, salts and prodrugs
thereof.
[0014] According to an aspect of the present invention, there is
provided a method of treating cancer comprising administering by
administering a PAR-4 agonist as part of a cancer treatment
regimen. In some instances the PAR-4 agonist is a cephalosporin
antibiotic, for instance a second generation cephalosporin
antibiotic such as cefuroxime, salts and prodrugs thereof. The
cancer treatment regimen can include administration of one or more
additional PAR-4 agonists, one or more additional chemotherapeutic
agents, exposure to ionizing radiation, and/or surgical
interventions.
[0015] According to an aspect of the present invention, there is
provided a method of treating cancer comprising administering
cefuroxime, or a salt or prodrug thereof, in combination with one
or more additional chemotherapeutic agents, in which the additional
chemotherapeutic agents are administered with cefuroxime either
simultaneously, sequentially, or separately.
[0016] The details of one or more embodiments are set forth in the
descriptions below. Other features, objects, and advantages will be
apparent from the description and from the claims.
BRIEF DESCRIPTION OF THE FIGURES
[0017] FIG. 1 depicts a Western blot analysis demonstrating that
cefuroxime causes increased PAR-4 secretion in mouse embryonic
fibroblast cells.
DETAILED DESCRIPTION
[0018] Before the present methods and systems are disclosed and
described, it is to be understood that the methods and systems are
not limited to specific synthetic methods, specific components, or
to particular compositions. It is also to be understood that the
terminology used herein is for the purpose of describing particular
embodiments only and is not intended to be limiting.
[0019] As used in the specification and the appended claims, the
singular forms "a," "an" and "the" include plural referents unless
the context clearly dictates otherwise. Ranges may be expressed
herein as from "about" one particular value, and/or to "about"
another particular value. When such a range is expressed, another
embodiment includes from the one particular value and/or to the
other particular value. Similarly, when values are expressed as
approximations, by use of the antecedent "about," it will be
understood that the particular value forms another embodiment. It
will be further understood that the endpoints of each of the ranges
are significant both in relation to the other endpoint, and
independently of the other endpoint.
[0020] "Optional" or "optionally" means that the subsequently
described event or circumstance may or may not occur, and that the
description includes instances where said event or circumstance
occurs and instances where it does not.
[0021] Throughout the description and claims of this specification,
the word "comprise" and variations of the word, such as
"comprising" and "comprises," means "including but not limited to,"
and is not intended to exclude, for example, other additives,
components, integers or steps. "Exemplary" means "an example of and
is not intended to convey an indication of a preferred or ideal
embodiment. "Such as" is not used in a restrictive sense, but for
explanatory purposes.
[0022] Disclosed are components that can be used to perform the
disclosed methods and systems. These and other components are
disclosed herein, and it is understood that when combinations,
subsets, interactions, groups, etc. of these components are
disclosed that while specific reference of each various individual
and collective combinations and permutation of these may not be
explicitly disclosed, each is specifically contemplated and
described herein, for all methods and systems. This applies to all
aspects of this application including, but not limited to, steps in
disclosed methods. Thus, if there are a variety of additional steps
that can be performed it is understood that each of these
additional steps can be performed with any specific embodiment or
combination of embodiments of the disclosed methods.
[0023] The compounds disclosed herein may be formulated as a
pharmaceutically acceptable salt. Pharmaceutically acceptable salts
are salts that retain the desired biological activity of the parent
compound and do not impart undesirable toxicological effects.
Examples of such salts are acid addition salts formed with
inorganic acids, for example, hydrochloric, hydrobromic, sulfuric,
phosphoric, and nitric acids and the like; salts formed with
organic acids such as acetic, oxalic, tartaric, succinic, maleic,
fumaric, gluconic, citric, malic, methanesulfonic,
ptoluenesulfonic, napthalenesulfonic, and polygalacturonic acids,
and the like; salts formed from elemental anions such as chloride,
bromide, and iodide; salts formed from metal hydroxides, for
example, sodium hydroxide, potassium hydroxide, calcium hydroxide,
lithium hydroxide, and magnesium hydroxide; salts formed from metal
carbonates, for example, sodium carbonate, potassium carbonate,
calcium carbonate, and magnesium carbonate; salts formed from metal
bicarbonates, for example, sodium bicarbonate and potassium
bicarbonate; salts formed from metal sulfates, for example, sodium
sulfate and potassium sulfate; and salts formed from metal
nitrates, for example, sodium nitrate and potassium nitrate.
Pharmaceutically acceptable and non-pharmaceutically acceptable
salts may be prepared using procedures well known in the art, for
example, by reacting a sufficiently basic compound such as an amine
with a suitable acid comprising a physiologically acceptable anion.
Alkali metal (for example, sodium, potassium, or lithium) or
alkaline earth metal (for example, calcium) salts of carboxylic
acids can also be made.
[0024] The cephalosporin antibiotics disclosed herein may be
formulated as pharmaceutically acceptable prodrugs. Typically
formed as ester with the 4-carboxylic acid group, prodrugs can
substantially increase the bioavailability of the compounds,
permitting more effective oral therapy. Such esters include
C.sub.1-C.sub.10 alkyl esters, which may or may not be substituted.
A preferred substituent is carbonyl-oxy and alkyloxy-carbonyloxy.
Exemplary esters include pivaloyloxy-methyl ester,
1-(isopropyloxy-carbonyloxy)ethyl ester, and 1-(acetyloxy)ethyl
ester.
[0025] Unchecked cell growth is a characteristic of all cancers.
Suppression of growth inhibitory or apoptotic functions by growth
stimulatory or cell survival proteins is seen in human cancer. The
coupling between cell division and cell death is thought to act as
a barrier that cells must overcome for cancer initiation and
progression. This may be the underlying reason why cancer cells
often over express anti-apoptotic proteins such as Bcl-2, Bcl-xL
and survivin, along with inactivation of pro-apoptotic
tumor-suppressor proteins p53, p19arf, PAR-4 and PTEN that control
apoptosis pathways, generating severe defects in the balance
between cell division and programmed cell death in cancer settings.
Thus the mentioned abnormalities that generate defects in apoptotic
pathways allow cancer cells to survive.
[0026] The inventors of the present invention have surprisingly
found that cefuroxime is a potent secretor or agonist of prostate
apoptosis response-4 (PAR-4). As such, cefuroxime can be used to
treat PAR-4 associated cancers, either alone or in combination with
other cancer therapy regimens.
[0027] The terms "induces, "secretor" or "agonist" are used
interchangeably throughout the specification, all the terms
indicate that the drug increases the expression of PAR-4.
[0028] The term "combination" as used herein, defines either a
fixed combination in one unit dosage form, a non-fixed combination
or a kit of parts for the combined administration.
[0029] The term "treating" or "treatment" as used herein comprises
a treatment relieving, reducing or alleviating at least one symptom
in a subject or effecting a delay of progression of a disease. For
example, treatment can be the diminishment of one or several
symptoms of a disorder or complete eradication of a disorder, such
as cancer. Within the meaning of the present invention, the term
"treat" also denotes to arrest, delay the onset (i.e., the period
prior to clinical manifestation of a disease) and/or reduce the
risk of developing or worsening a disease.
[0030] PAR-4 is a pro-apoptotic, tumor suppressor protein. It is
found to be deregulated in several cancers. Several studies have
documented the association of low level of PAR-4 with poor
prognosis in cancers of prostate, endometrial, renal, pancreas, and
breast. Endoplasmic reticulum-stress and higher levels of protein
kinase A in tumor cells confer the coveted feature of cancer
selective response to extracellular and intracellular PAR-4,
respectively. Recent studies have shown that systemic PAR-4 confers
resistance to tumor growth in mice.
[0031] PAR-4 is a leucine zipper domain protein identified in cells
undergoing apoptosis in response to exogenous insults. PAR-4 is
expressed ubiquitously among the various tissue types, and resides
in both the cytoplasm and the nucleus. Although endogenous PAR-4 is
largely inactivated, and does not produce extensive apoptosis by
itself, it is essential for the apoptotic function of diverse
cytotoxic agents. Interestingly, PAR-4 over-expression is
sufficient to induce apoptosis in most cancer cells, but not in
normal or immortalized cells.
[0032] PAR-4 has been shown to activate apoptosis through intrinsic
and extrinsic pathways. Upregulation or induction of PAR-4 by
apoptotic stimuli such as tumor necrosis factor alpha (TNF.alpha.),
TRAIL and Fas induce cell death in cancer cells. Other studies
showed that overexpression of PAR-4 enhances the activity of
anticancer drugs such as 5-fluorouracil and induces
radio-sensitivity. While the intracellular role of PAR-4 is
established and the mechanisms well studied, recent studies have
demonstrated that secretory or extracellular PAR-4 induces
apoptosis in cancer cells (Shastri et al., 2015).
[0033] Previous studies suggest that the role of PAR-4 in apoptosis
is cancer cell selective in that (i) overexpression of PAR-4
triggers apoptosis in various cancer cell lines but not in normal
and primary cells, (ii) depletion of PAR-4 by RNA interference
(RNAi) confers resistance in cancer cells, but not in primary
fibroblasts, to various apoptotic agents, and (iii) PAR-4 displays
proapoptotic functions in cells transformed with oncogene Ras but
not in normal cells. Recently, PAR-4 was shown to be secreted by
mammalian cells and, through interaction with the cell surface
receptor GRP78, to induce cancer cell apoptosis in a specific
manner.
[0034] PAR-4 is found to be down regulated in several cancers like
prostate, endometrial, renal, pancreas, and breast. Also because
the baseline levels of PAR-4 secreted by normal cells are generally
inadequate to cause massive apoptosis in cancer cell, secretogogues
that bolster the release of PAR-4 constitute an important
therapeutic advance. For example, Nutlin-3a, originally developed
as an MDM2 inhibitor, stimulated PAR-4 secretion at micromolar
levels in mouse embryonic fibroblast (MEF) cells.
[0035] The inventors of the present invention have surprisingly
found through various studies that cefuroxime, being an antibiotic,
has been found to increase the secretion of PAR-4 protein.
[0036] Cefuroxime has a chemical nomenclature of (6R,
7R)-3-{[(aminocarbonyl) oxy]
methyl}-7-{[(2Z)-2-(2-furyl)-2-(methoxyimino)
acetyl]amino}-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylic
acid and is chemically represented as--
##STR00001##
[0037] The term "cefuroxime" as per present invention is used in
broad sense to include not only "cefuroxime" per se but also its
pharmaceutically acceptable derivatives thereof. Suitable
pharmaceutically acceptable derivatives include pharmaceutically
acceptable salts like sodium, pharmaceutically acceptable solvates,
pharmaceutically acceptable hydrates, pharmaceutically acceptable
anhydrates, pharmaceutically acceptable enantiomers,
pharmaceutically acceptable esters, pharmaceutically acceptable
isomers, pharmaceutically acceptable polymorphs, pharmaceutically
acceptable prodrugs, pharmaceutically acceptable tautomers,
pharmaceutically acceptable complexes etc. Preferably, the
cefuroxime used to treat cancer is cefuroxime axetil (i.e.,
cefuroxime 1-(acetyloxy)ethyl ester).
[0038] The types of cancers which may be treated using PAR-4
agonists include: acute lymphoblastic leukemia (ALL), acute myeloid
leukemia (AML), cancer in adrenocortical carcinoma, adrenal cortex
cancer, AIDS-related cancers, Kaposi sarcoma, AIDS-related
lymphoma, primary CNS lymphoma, anal cancer, appendix cancer,
carcinoid tumors, astrocytomas, atypical teratoid/rhabdoid tumor,
basal cell carcinoma, skin cancer (nonmelanoma), bile duct cancer,
extrahepatic bladder cancer, bladder cancer, bone cancer (includes
Ewing sarcoma and osteosarcoma and malignant fibrous histiocytoma),
brain tumors, breast cancer, bronchial tumors, Burkitt lymphoma
(non-Hodgkin), carcinoid tumor, cardiac (heart) tumors, atypical
teratoid/rhabdoid tumor, embryonal tumors, germ cell tumors,
lymphoma, primary--cervical cancer, cholangiocarcinoma, chordoma,
chronic lymphocytic leukemia (CLL), chronic myelogenous leukemia
(CML), chronic myeloproliferative neoplasms, colorectal cancer,
colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma,
ductal carcinoma in situ (DCIS), embryonal tumors, central nervous
system, endometrial cancer, ependymoma, esophageal,
esthesioneuroblastoma, ewing sarcoma, extracranial germ cell tumor,
extragonadal germ cell tumor, eye cancer, intraocular melanoma,
retinoblastoma, fallopian tube cancer, fibrous histiocytoma of
bone, malignant, and osteosarcoma, gallbladder cancer, gastric
(stomach) cancer, gastric (stomach) cancer, gastrointestinal
carcinoid tumor, gastrointestinal stromal tumors (GIST),
gastrointestinal stromal tumors (GIST), germ cell tumors, central
nervous system, extracranial, extragonadal, ovarian testicular,
gestational trophoblastic disease, gliomas, hairy cell leukemia,
head and neck cancer, heart tumors, hepatocellular (liver) cancer,
histiocytosis, Langerhans Cell, Hodgkin's lymphoma, hypopharyngeal
cancer, intraocular melanoma, islet cell tumors, pancreatic
neuroendocrine tumors, Kaposi sarcoma, kidney--langerhans cell
histiocytosis, laryngeal cancer, laryngeal cancer and
papillomatosis, leukemia, lip and oral cavity cancer, liver cancer
(primary), lung cancer, lung cancer, lymphoma-macroglobulinemia,
Waldenstrom--Non-Hodgkin lymphoma, male breast cancer, malignant
fibrous histiocytoma of bone and osteosarcoma, melanoma,
intraocular (eye), Merkel cell carcinoma, mesothelioma, malignant,
mesothelioma, metastatic squamous neck cancer with occult primary,
midline tract carcinoma involving NUT gene, mouth cancer, multiple
endocrine neoplasia syndromes, multiple myeloma/plasma cell
neoplasms, mycosis fungoides, myelodysplastic syndromes,
myelodysplastic/myeloproliferative neoplasms and chronic
myeloproliferative neoplasms, myelogenous leukemia, chronic (CML),
myeloid leukemia, acute (AML), nasal cavity and paranasal sinus
cancer, nasopharyngeal cancer, nasopharyngeal cancer,
neuroblastoma, non-hodgkin lymphoma, non-small cell lung cancer,
oral cancer, lip and oral cavity cancer and oropharyngeal cancer,
osteosarcoma and malignant fibrous histiocytoma of bone, ovarian
cancer, pancreatic cancer and pancreatic neuroendocrine tumors
(islet cell tumors), papillomatosis, paraganglioma, paraganglioma,
paranasal sinus and nasal cavity cancer, parathyroid cancer, penile
cancer, pharyngeal cancer, pheochromocytoma, pheochromocytoma,
pituitary tumor, plasma cell neoplasm/multiple myeloma,
pleuropulmonary blastoma, pregnancy and breast cancer, primary
central nervous system (CNS) lymphoma, primary peritoneal cancer,
prostate cancer, rectal cancer, renal cell (kidney) cancer,
retinoblastoma, rhabdomyosarcoma, salivary gland cancer, salivary
gland tumors, Ewing sarcoma, Kaposi sarcoma, osteosarcoma,
rhabdomyosarcoma, uterine sarcoma, vascular tumors, Sezary
syndrome, skin cancer, small cell lung cancer, small intestine
cancer, soft tissue sarcoma, squamous cell carcinoma, squamous neck
cancer with occult primary, metastatic, stomach (gastric) cancer,
stomach (gastric) cancer, T-cell lymphoma, cutaneous, testicular
cancer, throat cancer, thymoma and thymic carcinoma, thyroid
cancer, ureter and renal pelvis, transitional cell cancer, urethral
cancer, uterine cancer, endometrial and uterine sarcoma, vaginal
cancer, vaginal cancer, vascular tumors, vulvar cancer, Waldenstrom
Macroglobulinemia, Wilms Tumor.
[0039] In certain preferred embodiments, cefuroxime can be used to
treat prostate cancer, breast cancer, lung cancer, or skin
cancer/melanoma. In particular, cefuroxime can be used to treat
superficial spreading melanoma, nodular melanoma, lentigno maligna
melanoma, and desmoplastic melanoma. In some embodiments, the
patient to be treated does not have renal cancer.
[0040] Depending on the pathological stage, patient's age and other
physiological parameters, size of the tumor, and the extent of
invasion, the pharmaceutical composition comprising cefuroxime may
require specific dosage amounts and specific frequency of
administrations. Preferably, cefuroxime may be administered at
least once or multiple times a day in the dosage range from about 1
mg/kg/per day to about 100 mg/kg/per day. In some embodiments,
cefuroxime can be administered once, twice, three times, or greater
than three times a day, for a period of 1 week, 2 weeks, 4 weeks, 8
weeks, 12 weeks, 16 weeks, 20 weeks, 24 weeks, 28 weeks, 32 weeks,
42 weeks or 52 weeks. In some instances, cefuroxime can be
administered intermittently, for instance for a period of 1-10
days, followed by a period in which no cefuroxime is administered
(e.g., 1-10 days), followed by another period e.g., 1-10 days, in
which cefuroxime is administered. The on/off dosing schedule can be
repeated as many times as necessary.
[0041] In certain embodiments, cefuroxime can be administered in
combination with one or more chemotherapeutic agents. Exemplary
agents include Abiraterone, Methotrexate, Abraxane (Paclitaxel
Albumin-stabilized Nanoparticle Formulation, Brentuximab Vedotin,
Ado-Trastuzumab, Emtansine, Adriamycin, Afatinib, Everolimus,
Akynzeo (Netupitant and Palonosetron Hydrochloride), Imiquimod,
Aldesleukin, Alectinib, Alemtuzumab, Melphalan, Pemetrexed,
Palonosetron, Chlorambucil, Aminolevulinic Acid, Anastrozole,
Aprepitant, Pamidronate, Anastrozole, Exemestane, Nelarabine,
Arsenic Trioxide, Ofatumumab, Asparaginase Erwinia chrysanthemi,
Atezolizumab, Bevacizumab, Axitinib, Azacitidine, BEACOPP,
Carmustine, Belinostat, Bendamustine, BEP, Bevacizumab, Bexarotene,
Tositumomab, Bicalutamide, Carmustine, Bleomycin, Blinatumomab,
Bortezomib, Bosutinib, Brentuximab Vedotin, Busulfan, Cabazitaxel,
Cabozantinib-Alemtuzumab), Irinotecan, Capecitabine, CAPDX,
Fluorouracil, Carboplatin, carboplatin-taxol, Carfilzomib,
Carmustine, Bicalutamide, Lomustine, CEM, Ceritinib, Daunorubicin,
Cervarix (Recombinant HPV Bivalent Vaccine), Cetuximab,
Chlorambucil, CHOP, Cisplatin, Clofarabine, CMF, Cobimetinib,
Cabozantinib, COPDAC, COPP, COPP-ABV, Dactinomycin, Cobimetinib,
Crizotinib, CVP, Ifosfamide, Ramucirumab, Cytarabine,
Cyclophosphamide, Dabrafenib, Dacarbazine, Decitabine,
Dactinomycin, Daratumumab, Dasatinib, Daunorubicin, efibrotide
Sodium, Defibrotide, Degarelix, Denileukin Diftitox, Denosumab,
DepoCyt (Cytarabine Liposome), Dexamethasone, Dexrazoxane
Dinutuximab, Docetaxel, Doxorubicin, Dacarbazine, Rasburicase,
Epirubicin, Elotuzumab, Oxaliplatin, Eltrombopag Olamine,
Aprepitant, Empliciti (Elotuzumab), Enzalutamide, Epirubicin,
EPOCH, Cetuximab, Eribulin, Vismodegib, Erlotinib, Erwinaze
(Asparaginase, rwinia chrysanthemi), Etopophos (Etoposide
Phosphate), Etoposide, Etoposide Phosphate, Everolimus, Evista
(Raloxifene Hydrochloride), Evomela (Melphalan Hydrochloride),
Exemestane, 5-FU, Fareston (Toremifene), Farydak (Panobinostat),
Faslodex (Fulvestrant), FEC, Letrozole, Filgrastim, Fludarabine,
Flutamide, Methotrexate, Folfiri, Folfiri-Bevacizumab,
Folfiri-Cetuximab, Folfirinox, Folfox, Pralatrexate, FU-LV,
Fulvestrant, Gardasil (Recombinant HPV Quadrivalent Vaccine),
Obinutuzumab, Gefitinib, Gemcitabine, Gemtuzumab Ozogamicin,
Afatinib, Imatinib, Carmustine, Glucarpidase, Goserelin Acetate,
Eribulin, Trastuzumab, Topotecan ,HydroxyureaPalbociclib),
Ibritumomab Tiuxetan, Ibrutinib, ICE, Ponatinib, Idarubicin,
Idelalisib, Ifex (Ifosfamide), Ifosfamide, IL-2 (Aldesleukin),
Imatinib Ibrutinib, Imiquimod, Imlygic, Talimogene Laherparepvec,
Axitinib, Interferon Alfa-2b, Recombinant, Interleukin-2 ,
Aldesleukin), Intron A (Recombinant Interferon Alfa-2b),
Tositumomab, Ipilimumab, Gefitinib, Irinotecan, Romidepsin,
Ixabepilone, Ixazomib, Ruxolitinib, Cabazitaxel, Ado-Trastuzumab
Emtansine), Raloxifene, Palifermin, Pembrolizumab, Carfilzomib ,
Lanreotide, Lapatinib, Lenalidomide Lenvatinib, Letrozole,
Leucovorin, Leukeran , Chlorambucil), Leuprolide, Olaparib,
Vincristine, Procarbazine, Mechlorethamine, Megestrol, Trametinib,
Melphalan, Mercaptopurine, Mesna, Temozolomide, Methotrexate,
Mitomycin C, Mitoxantrone, MOPP, Plerixafor, Mechlorethamine,
Busulfan, Azacitidine, Gemtuzumab Ozogamicin, Vinorelbine,
Necitumumab, Nelarabine, Neosar (Cyclophosphamide), Netupitant and
Palonosetron Hydrochloride, Neupogen (Filgrastim), Nexavar
(Sorafenib Tosylate), Nilotinib, Ninlaro (Ixazomib Citrate),
Nivolumab, Nolvadex (Tamoxifen Citrate), Nplate (Romiplostim),
Obinutuzumab, Odomzo (Sonidegib), OEPA, Ofatumumab, Olaparib,
Omacetaxine Mepesuccinate, Oncaspar (Pegaspargase), Ondansetron
Hydrochloride, Onivyde (Irinotecan Hydrochloride Liposome), Ontak
(Denileukin Diftitox), Opdivo (Nivolumab), OPPA, Osimertinib,
Oxaliplatin, Paclitaxel, Paclitaxel Albumin-stabilized Nanoparticle
Formulation, Palbociclib, Palifermin, Palonosetron Hydrochloride,
Palonosetron Hydrochloride and Netupitant, Pamidronate Di sodium,
Panitumumab, Panobinostat, Paraplat (Carboplatin), Paraplatin
(Carboplatin), Pazopanib Hydrochloride, Pegaspargase, Peginterferon
Alfa-2b, PEG-Intron (Peginterferon Alfa-2b), Pembrolizumab,
Pemetrexed Disodium, Perjeta (Pertuzumab), Pertuzumab, Platinol
(Cisplatin), Platinol-AQ (Cisplatin), Plerixafor, Pomalidomide,
Pomalyst (Pomalidomide), Ponatinib Hydrochloride, Portrazza
(Necitumumab), Pralatrexate, Prednisone, Procarbazine
Hydrochloride, Proleukin (Aldesleukin), Prolia, Denosumab),
Promacta (Eltrombopag Olamine), Provenge (Sipuleucel-T), Purinethol
(Mercaptopurine), Purixan (Mercaptopurine), Radium 223 Dichloride,
Raloxifene Hydrochloride, Ramucirumab, Rasburicase, R-CHOP, R-CVP,
Recombinant Human Papillomavirus (HPV) Bivalent Vaccine,
Recombinant Human Papillomavirus (HPV) Nonavalent Vaccine,
Recombinant Human Papillomavirus (HPV) Quadrivalent Vaccine,
Recombinant Interferon Alfa-2b, Regorafenib, R-EPOCH, Revlimid
(Lenalidomide), Rheumatrex (Methotrexate), Rituxan , ituximab),
Rituximab, Rolapitant Hydrochloride, Romidepsin, Romiplostim,
Rubidomycin , Daunorubicin Hydrochloride), Ruxolitinib Phosphate,
Sclerosol Intrapleural Aerosol (Talc), Siltuximab, Sipuleucel-T,
Somatuline Depot (Lanreotide Acetate), Sonidegib, Sorafenib
Tosylate, Sprycel (Dasatinib), STANFORD V, Sterile Talc Powder
(Talc), Steritalc (Talc), Stivarga (Regorafenib), Sunitinib Malate,
Sutent (Sunitinib Malate), Sylatron (Peginterferon Alfa-2b),
Sylvant (Siltuximab), Synovir (Thalidomide), Synribo (Omacetaxine
Mepesuccinate), Tabloid (Thioguanine), TAC, Tafinlar (Dabrafenib),
Tagrisso (Osimertinib), Talimogene Laherparepvec, Tamoxifen
Citrate, Tarabine PFS (Cytarabine), Tarceva (Erlotinib
Hydrochloride), Targretin , Bexarotene), Tasigna (Nilotinib), Taxol
(Paclitaxel), Taxotere (Docetaxel), Tecentriq ,
Atezolizumab),Temodar (Temozolomide), Temozolomide, Temsirolimus,
Thalidomide, Thalomid (Thalidomide), Thioguanine, Thiotepa, Tolak
(Fluorouracil--Topical), Topotecan Hydrochloride, Toremifene,
Torisel (Temsirolimus), Tositumomab and Iodine I 131 Tositumomab,
Totect (Dexrazoxane Hydrochloride), Trabectedin, Trametinib,
Trastuzumab, Treanda , Bendamustine Hydrochloride), Trifluridine
and Tipiracil Hydrochloride, Trisenox (Arsenic Trioxide), Tykerb
(Lapatinib Ditosylate), Unituxin (Dinutuximab), Uridine Triacetate,
Vandetanib, VAMP, Varubi (Rolapitant Hydrochloride), Vectibix
(Panitumumab), Velban (Vinblastine Sulfate), Velcade (Bortezomib),
Velsar (Vinblastine Sulfate), Vemurafenib, Venclexta (Venetoclax),
Venetoclax, Viadur (Leuprolide Acetate), Vidaza (Azacitidine),
Vinblastine Sulfate, Vincasar PFS (Vincristine Sulfate),
Vincristine Sulfate, Vincristine Sulfate Liposome, Vinorelbine
Tartrate, Vismodegib, Vistogard (Uridine Triacetate), Voraxaze ,
Glucarpidase), Vorinostat, Votrient (Pazopanib Hydrochloride),
Wellcovorin (Leucovorin Calcium), Xalkori (Crizotinib), Xeloda
(Capecitabine), XELIRI, XELOX, Xgeva (Denosumab), Xofigo (Radium
223 Dichloride), Xtandi (Enzalutamide), Yervoy (Ipilimumab),
Yondelis (Trabectedin), Zaltrap (Ziv-Aflibercept), Zarxio
(Filgrastim), Zelboraf (Vemurafenib), Zevalin (Ibritumomab
Tiuxetan), Zinecard (Dexrazoxane Hydrochloride), Ziv-Aflibercept,
Zofran (Ondansetron Hydrochloride), Zoladex (Goserelin Acetate),
Zoledronic Acid, Zolinza (Vorinostat), Zometa (Zoledronic Acid),
Zydelig (Idelalisib), Zykadia (Ceritinib), Zytiga (Abiraterone
Acetate). Preferred additional agents include tamoxifen,
bendamustine, cladribine, or a combination thereof.
[0042] Cefuroxime can be administered with one or more
chemotherapeutic agents either simultaneously, sequentially, or
separately. In certain cases, cefuroxime can be administered for a
period of at least 1 week, at least 2 weeks, at least 4 week, at
least 6 weeks, at least 8 week, or at least 10 weeks, prior to
commencing treatment with additional chemotherapeutic agents. In
some instances, cefuroxime and the other agent can be administered
intermittently, for instance a period of cefuroxime administration,
followed by a period in which the other agent to administered,
followed by another period of cefuroxime administration. The cycle
can be repeated as many times as necessary.
[0043] In certain cases, the combination of cefuroxime and
additional agent will exhibit a greater than additive effect (i.e.,
a synergistic effect). In other instance, the use of cefuroxime
permits a reduced amount of the other agent to be administered,
without a corresponding decrease in therapeutic efficiency.
[0044] In some instances, cefuroxime can be used in combination
with ionizing radiation and/or surgical interventions for the
treatment of cancer. Cefuroxime can be administered before, during,
or after treatment with ionizing radiation or surgical
intervention. In certain cases, cefuroxime can be administered for
a period of at least 1 week, at least 2 weeks, at least 4 week, at
least 6 weeks, at least 8 week, or at least 10 weeks, prior to
commencing treatment with ionizing radiation or surgery. Exemplary
forms of radiation include x-rays, gamma rays, electron beams and
proton beams. It has been found that administration of cefuroxime
permits a reduction in the total exposure of the patient to
ionizing radiation, without a corresponding reduction in
therapeutic efficiency. In certain instances, cefuroxime can be
administered both prior and subsequent to ionizing radiation and/or
surgical interventions.
[0045] The use of cefuroxime may preferably be associated with one
or more of the above referenced anti-cancer drugs as a combination
therapy (either of the same functional class or other) depending on
various factors like drug-drug compatibility, patient compliance
and other such factors wherein the said combination therapy may be
administered either simultaneously, sequentially, or separately for
the treatment of cancer.
[0046] It may be well appreciated by a person skilled in the art
that the pharmaceutical composition comprising cefuroxime in
combination with one or more anti-cancer drugs may require specific
dosage amounts and specific frequency of administrations
specifically considering their individual established doses, the
dosing frequency, patient adherence and the regimen adopted. As
described herein, considering that there are various parameters to
govern the dosage and administration of the combination composition
as per the present invention, it would be well acknowledged by a
person skilled in the art to exercise caution with respect to the
dosage, specifically, for special populations associated with other
disorders.
[0047] Preferably, cefuroxime may be provided in the form of a
pharmaceutical composition such as but not limited to, unit dosage
forms including tablets, capsules (filled with powders, pellets,
beads, mini-tablets, pills, micro-pellets, small tablet units,
multiple unit pellet systems (MUPS), disintegrating tablets,
dispersible tablets, granules, and microspheres,
multiparticulates), sachets (filled with powders, pellets, beads,
mini-tablets, pills, micro-pellets, small tablet units, MUPS,
disintegrating tablets, dispersible tablets, granules, and
microspheres, multiparticulates), powders for reconstitution,
transdermal patches and sprinkles, however, other dosage forms such
as controlled release formulations, lyophilized formulations,
modified release formulations, delayed release formulations,
extended release formulations, pulsatile release formulations, dual
release formulations and the like. Liquid or semisolid dosage form
(liquids, suspensions, solutions, dispersions, ointments, creams,
emulsions, microemulsions, sprays, patches, spot-on), injection
preparations, parenteral, topical, inhalations, buccal, nasal etc.
may also be envisaged under the ambit of the invention. The
pharmaceutical composition may further include one or more
additional therapeutic agents as described above. In other
instances, cefuroxime may be provided as a component in a kit that
also contains one or more additional therapeutic agents, or
instructions for administering cefuroxime to a patient in need
thereof.
[0048] The inventors of the present invention have also found that
the solubility properties of cefuroxime may be improved by
nanosizing thus leading to better bioavailability and dose
reduction of the drug.
[0049] In one embodiment, cefuroxime may be present in the form of
nanoparticles which have an average particle size of less than 2000
nm.
[0050] Suitable excipients may be used for formulating the dosage
forms according to the present invention such as, but not limited
to, surface stabilizers or surfactants, viscosity modifying agents,
polymers including extended release polymers, stabilizers,
disintegrants or super disintegrants, diluents, plasticizers,
binders, glidants, lubricants, sweeteners, flavoring agents,
anti-caking agents, opacifiers, anti-microbial agents, antifoaming
agents, emulsifiers, buffering agents, coloring agents, carriers,
fillers, anti-adherents, solvents, taste-masking agents,
preservatives, antioxidants, texture enhancers, channeling agents,
coating agents or combinations thereof.
EXAMPLES
[0051] The following examples are set forth below to illustrate the
methods and results according to the disclosed subject matter.
These examples are not intended to be inclusive of all aspects of
the subject matter disclosed herein, but rather to illustrate
representative methods, compositions, and results. These examples
are not intended to exclude equivalents and variations of the
present invention, which are apparent to one skilled in the
art.
[0052] Efforts have been made to ensure accuracy with respect to
numbers (e.g., amounts, temperature, etc.) but some errors and
deviations should be accounted for. Unless indicated otherwise,
parts are parts by weight, temperature is in .degree. C. or is at
ambient temperature, and pressure is at or near atmospheric. There
are numerous variations and combinations of reaction conditions,
e.g., component concentrations, temperatures, pressures, and other
reaction ranges and conditions that can be used to optimize the
product purity and yield obtained from the described process. Only
reasonable and routine experimentation will be required to optimize
such process conditions.
Example 1
Cefuroxime induced PAR-4 Secretion
[0053] A. Assay Protocol to Analyse Non-Cytotoxic Concentration
[0054] Mouse embryonic fibroblast cells were plated in 96-well cell
culture plates (2.times.10.sup.4 cells/well) and were treated with
drug/s at various concentrations. Treated cells were maintained at
37.degree. C. in 5% CO.sub.2 for 96 hrs. After the incubation
anti-proliferative activity of test compounds was measured using
ProMega Cell Titer aqueous one solution cell proliferation assay
kit, viz. CellTiter 96.RTM. AQueous One Solution Reagent. Cells
were incubated for 1-4 hours at 37.degree. C., 5% CO.sub.2
incubator and absorbance was measured at 490 nM using a plate
reader. IC.sub.50 values were determined by plotting compound
concentration versus cell viability.
[0055] B. Assay Protocol to Analyze PAR-4 Secretion
[0056] Material: DMEM media, FBS, Mouse embryonic fibroblast cells
, PAR-4 antibody, Antirabbit IgG-HRP, Collagen Al antibody, B-actin
antibody, Antimouse IgG HRP
[0057] Day 1: Cell Seeding
[0058] Six well plate seeded (2 ml per well) with Mouse embryonic
fibroblast cells in DMEM +10% FBS. The plate was incubated for
16-18 hrs, 37.degree. C. 5% CO.sub.2.
[0059] Day 2: Drug Addition
[0060] Drugs to be tested are diluted with suitable solvent to
obtain 25 mM concentration. Media from cells is discarded and
replaced with DMEM media with 0.1%FBS. Compound added to the cells
at final concentration of 25 .mu.M.
[0061] Day 3: Sample Preparation
[0062] Conditioned media (CM-supernatant) is collected from each
well and centrifuged at 1000 rpm/5 mins to remove cell debris.
After centrifugation the media was poured in concentration filters
(Amicon ultracentrifuge tube with cutoff 10 Kda) and again
centrifuged at 4500 rpm/15 mins at 4.degree. C. The media was
concentrated to 100 .mu.l and then mixed with 50 .mu.l of loading
dye. The mixture was maintained in a 100.degree. C. dry bath/water
bath for 10 mins. SDS-PAGE and a western blot was performed of the
samples using broad range molecular weight marker.
[0063] C. In Vitro Cytotoxicity on Cancer Cells
[0064] Various cancer cells lines were plated in 96-well cell
culture plates (2.times.10.sup.4 cells/well) and were treated with
drug/s at various concentrations. Treated cells were maintained at
37.degree. C. in 5% CO.sub.2 for 96 hrs. After the incubation the
antiproliferative activity of test compounds was measured using
ProMega Cell Titer aqueous one solution cell proliferation assay
kit, viz. CellTiter 96.RTM. Aqueous One Solution Reagent. Cells
were incubated for 1-4 hours at 37.degree. C., 5% CO.sub.2
incubator and absorbance was measured at 490 nM using a plate
reader. IC.sub.50 values were determined by plotting compound
concentration versus cell viability.
[0065] Results [0066] A. Non-Cytotoxic concentration determination:
Cefuroxime does not show toxicity in Mouse Embryonic fibroblast
cells up to 48 hours at the highest concentration of 125 .mu.M.
[0067] B. PAR-4 Secretion: [0068] 1. Cefuroxime exhibited 4-5-fold
increase in the secretion of PAR-4 in the conditioned media from
the mouse embryonic fibroblast cells when compared to DMSO control
as studied by Western blot analysis. [0069] 2. Cefuroxime caused
about 2 fold increase in the secretion of intracellular PAR-4 from
the mouse embryonic fibroblast cells when compared to DMSO control
as studied by Western blot analysis. [0070] 3. Albumin and collagen
was used as a loading control for conditioned media. [0071] 4.
Actin was used as loading control for lysate. [0072] 5. Also the
conditioned media was cytotoxic to various panel of cancer cell
lines.
TABLE-US-00001 [0072] Cefuroxime Axetil for Oral Suspension
Ingredients Qty. mg/g Cefuroxime axetil 20-500 Polysorbate 80
0.25-0.50 Simethicone 0.6-1.0 Xanthan gum 10-20 Silicon dioxide
7.5-12.5 Titanium dioxide 15-20 Sodium benzoate 6-10 Cherry flavor,
natural and 2.5-5.0 artificial (microencapsulated) Sucrose q.s.t.
1000 mg
[0073] Cefuroxime axetil, xanthan gum, silicon dioxide, titanium
dioxide, sodium benzoate, cherry flavor and a portion of sucrose
were sifted. Polysorbate 80 and Simethicone were combined with a
portion of sucrose and sifted. The two mixtures were then blended.
The resulting blend was filled in white translucent HDPE bottle
with cap and sealed using induction sealer.
TABLE-US-00002 Cefuroxime axetil suspension Ingredients Quantity
mg/mL Cefuroxime axetil 20-500 Sorbitol solution 70% 30-300
Saccharin 15-30 Cherry flavor 2.5-5 Water purified q.s.t. 1 mL
[0074] Cefuroxime axetil was added to the sorbitol solution and
mixed vigorously to form a suspension. Saccharin and cherry flavor
were dissolved in purified water and added to the suspension. The
desired volume was achieved by addition of purified water.
TABLE-US-00003 Cefuroxime axetil tablets Ingredients Quantity
mg/tablet Cefuroxime axetil 20-750 Lactose monohydrate 30-250
Microcrystalline cellulose 40-300 (Avicel PH 101) Pregelatinized
starch 30-90 Croscarmellose sodium 15-45 Poloxamer 188 (Pulmonic F
68) 5-20 Silicon dioxide colloidal 2.5-10 Magnesium stearate 3-10
Purified water q.s Coating Opadry ready mix 10-25 Purified water
qs
[0075] Cefuroxime axetil, lactose monohydrate, microcrystalline
cellulose, pregelatinized starch, and a portion (one-half) of
croscarmellose sodium were sifted. The sifted powders were loaded
in a suitable mixer/granulator and mixed. Poloxamer 188 was
dissolved in a sufficient quantity of purified water, and the
solution was used to wet granulate the mixed powder. Granules were
dried and then blended with pre-sifted silicon dioxide,
microcrystalline cellulose, and croscarmellose sodium. The blend
was lubricated using pre-sifted magnesium stearate, compressed into
tablets and coating with opadry ready mix.
TABLE-US-00004 Cefuroxime for Injection Ingredients Qty/vial (mg)
Cefuroxime sodium 20-750 Dextrose hydrous, USP 40-1500 Sodium
citrate hydrous 250-500 Hydrochloric acid for pH adjustment q.s
Sodium hydroxide for pH adjustment q.s Water for injection, USP Qst
2 mL
[0076] Dextrose hydrous and sodium citrate hydrous were dissolved
in freshly distilled water. Cefuroxime sodium was added and stirred
to dissolve. The pH of the solution was adjusted to 5.0-7.5 using
hydrochloric acid or sodium hydroxide and volume was made up with
water. The solution of step 3 was filtered through a 0.22-.mu.m
filter and filled aseptically into vials and freeze dried.
[0077] The compositions and methods of the appended claims are not
limited in scope by the specific compositions and methods described
herein, which are intended as illustrations of a few aspects of the
claims and any compositions and methods that are functionally
equivalent are intended to fall within the scope of the claims.
Various modifications of the compositions and methods in addition
to those shown and described herein are intended to fall within the
scope of the appended claims. Further, while only certain
representative compositions and method steps disclosed herein are
specifically described, other combinations of the compositions and
method steps also are intended to fall within the scope of the
appended claims, even if not specifically recited. Thus, a
combination of steps, elements, components, or constituents may be
explicitly mentioned herein or less, however, other combinations of
steps, elements, components, and constituents are included, even
though not explicitly stated. The term "comprising" and variations
thereof as used herein is used synonymously with the term
"including" and variations thereof and are open, non-limiting
terms. Although the terms "comprising" and "including" have been
used herein to describe various embodiments, the terms "consisting
essentially of and "consisting of can be used in place of
"comprising" and "including" to provide for more specific
embodiments of the invention and are also disclosed. Other than in
the examples, or where otherwise noted, all numbers expressing
quantities of ingredients, reaction conditions, and so forth used
in the specification and claims are to be understood at the very
least, and not as an attempt to limit the application of the
doctrine of equivalents to the scope of the claims, to be construed
in light of the number of significant digits and ordinary rounding
approaches.
* * * * *