Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Status of the Claims
Per Applicant’s amendment to the claims, submitted on 08/08/2024, claims 1-18 are amended, and claims 19-20 are canceled. Currently, claims 1-18 are pending in the instant application.
Information Disclosure Statement
The information disclosure statement (IDS) submitted on 08/08/2024 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
Specification
The disclosure is objected to because it contains an embedded hyperlink and/or other form of browser-executable code. Applicant is required to delete the embedded hyperlink and/or other form of browser-executable code; references to websites should be limited to the top-level domain name without any prefix such as http:// or other browser-executable code. See MPEP § 608.01.
Browser executable code is found on page 1 of the specification in the following section:
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Claim Rejections - 35 USC § 112 – Second Paragraph
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1, 4, 11, and 14 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 1 is indefinite for reciting the phrase “Urolithin or a precursor thereof” (underlined for emphasis), because a person of ordinary skill in the art would not reasonably be able to understand the metes and bounds of the claim. Under broadest reasonable interpretation, a “precursor” may encompass any chemical compound or component that can be used to form urolithin. Furthermore, urolithin exists in multiple forms (i.e., A, B, C, D, etc.). It is unclear what compounds or substances the instant claim is limited to in regards to “precursors”.
Claim 4 is indefinite for reciting the term “preferably”, because a person of ordinary skill in the art would not reasonably be able to understand the metes and bounds of the claim. The indicated recitation is indefinite because it is unclear whether the limitation(s) following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Claim 11 is indefinite for reciting the phrase “wherein the preceding cancer therapy involves use of a urolithin”, because a person of ordinary skill in the art would not reasonably be able to understand the metes and bounds of the claim. In particular, the recitation of a therapy that “involves use of urolithin” is indefinite because it is unclear as to what the recited use entails. This rejection may be overcome by amending the claim to clarify how urolithin is involved with the preceding therapy (i.e., administration of urolithin, etc.).
Claim 14 is indefinite for reciting the phrase “urolithin or precursor thereof”, because a person of ordinary skill in the art would not reasonably be able to understand the metes and bounds of the claim. Under broadest reasonable interpretation, a “precursor” may encompass any chemical compound or component that can be used to form urolithin. Furthermore, urolithin exists in multiple forms (i.e., A, B, C, D, etc.). It is unclear what compounds or substances the instant claim is limited to in regards to a “precursor thereof”.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claim(s) 1 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez (Food and Chemical Toxicology 92 (2016) 8-16).
Claim 1 recites a method for use in reducing the risk of the recurrence of cancer or tumor formation in a patient in need thereof in the remission phase of a preceding cancer therapy in said patient comprising administering a composition comprising a therapeutically effective amount of a Urolithin or a precursor thereof to a patient in need of the same.
Sanchez teaches the treatment of cancer by administering compositions comprising urolithins and ellagic acid. Sanchez does not explicitly teach the administration of said compositions to a patient in the remission phase of a cancer therapy. However, it would be obvious to administer to such a patient because Sanchez indicates use in the prevention of cancer recurrence, and there would be a reasonable expectation of success in preventing such recurrence in a patient in cancer remission.
Sanchez indicates that compositions comprising urolithin A, urolithin C, and ellagic acid (EA) were capable of reducing phenotypic and molecular features in colon cancer stem cell lines of Caco-2 and primary tumors of colorectal cancer (CRC) (page 8, Abstract)1. Two distinct urolithin mixtures were tested: (i) MPhA comprising 85% Uro-A, 10% Uro-C, 5% EA; and (ii) MPhB comprising 30% Uro-A, 50% IsoUro-A,10% Uro-B, 5% Uro-C, 5% EA. Both MPhA and MPhB were dosed to cell lines in a low concentration (C1) and a high concentration (C2). Dosing to the Caco-2 cell line provided the following results (page 11, Fig 1):
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As can be seen above, at least with regards to the C2 high dose, both MPhA and MPhB compositions provided a decrease in colonosphere number (A) and a decrease in average colonosphere size (B and C). MPhA was further capable of significantly decreasing ALDH activity (D).
Dosing to primary tumor CRC lines yielded the following results (page 12, Fig 2):
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As can be seen above, MPhA was capable of significantly reducing number of colonospheres (A) and ALDH activity (D) at the C2 high dose. Both MPhA and MPhB were capable of reducing colonosphere size (B and C). The results of both assays appear to indicate that the compositions, especially MPhA, have inhibitory action on colon CSC activity (page 13)2.
While Sanchez does not explicitly teach the administration of their compositions to patients in cancer remission, they do indicate that the compositions disclosed are useful for the prevention of cancer recurrence (page 9)3. Sanchez further suggests the oral intake of such compounds to combat colon CSCs in vivo (page 13)4. As the teachings of Sanchez are relevant to not only the treatment of cancer, but the recurrence of cancer, it would be obvious for a person of ordinary skill to contemplate applying such treatment to patients in cancer remission. By definition, a patient in the post-treatment cancer remission phase would be a person having undergone cancer therapy, and having cancer symptoms either eliminated or lessened. Such a patient would be at risk for cancer recurrence.
Accordingly, given the teachings of Sanchez, a person of ordinary skill in the art would have found it prima facie obvious to administer a urolithin composition to a patient in cancer remission, as there would be a reasonable expectation of success in preventing cancer recurrence in said patient.
Claim 2 further limits the method of claim 1 wherein the patient is a human or non-human mammal.
While Sanchez does not explicitly teach in-vivo administration of their compositions to a human or non-human mammal, they do teach administration ex-vivo to human harvested colon tumor cells. Accordingly, administration to at least a human subject would be obvious to a person of ordinary skill in the art.
Claim(s) 3 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez in view of Vicinanza (Evidence-Based Complementary and Alternative Medicine Volume 2013, Article ID 247504).
Claim 3 further limits the method of claim 1 wherein the patient is an immune compromised patient.
As discussed previously, Sanchez teaches a method of reducing cancer recurrence by administration of a composition comprising urolithins. Sanchez does not explicitly teach administration to an immune-compromised patient. However, it would be obvious to administer said composition to such a patient because Vicinanza teaches the administration of urolithins and ellagic acid to cancer cell lines extracted from immunodeficient mice, and there would be a reasonable expectation of success in treating cancer in immunodeficient patients.
Vicinanza provides an overview of the anti-cancer effects of pomegranate juice metabolites. More specifically, Vicinanza indicates that etalligitannins from pomegranate juice are metabolized by the body to form ellagic acid and urolithin A (page 1, Abstract)5, which provided dose dependent antiproliferative effects in DU-145 and PC-3 prostate cancer cell lines (page 4)6. The results are exemplified in the following graphs (page 5, Fig. 2):
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As can be seen in the graphs above, both ellagic acid and urolithin A were capable of reducing proliferation in both cell lines in both a dose and time dependent manner. Vicinanza further indicates that in-vitro testing was conducted on tumor tissue extracted from castrated severe combined immunodeficiency (SCID) mice with Los Angeles Prostate Cancer-4 (LAPC-4) human PCa xenografts, and that said testing revealed that administration of pomegranate extract was able to inhibit PCa proliferation (page 2)7. Given the teachings of Vicinanza, there would have been a reasonable expectation that administering a combination of ellagic acid and urolithin A would have been effective in reducing risk cancer recurrence in a patient having compromised immune system.
In summary, it would have been prima facie obvious at the time of invention for a person of ordinary skill in the art to be able to develop a method of the instant claim by combining the teachings of Sanchez and Vicinanza. Because Sanchez teaches a method of reducing risk of cancer recurrence by administering a combination of ellagic acid and urolithins, and Vicinanza teaches the use the same components (in pomegranate extract) for cancer treatment as applied to a cell line sourced from immune-compromised subjects, there would have been a reasonable expectation of success in reducing cancer recurrence in immune-compromised patients.
Claim(s) 4-6 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez.
Claim 4 further limits the method of claim 1 wherein the Urolithin or a precursor thereof is selected from the group consisting of a urolithin, ellagic acid, and an ellagitannin, more preferably is selected from Urolithin A, Urolithin B, Urolithin C or Urolithin D, er-and a combination thereof.
As discussed previously, Sanchez teaches administration of compositions comprising urolithin A, urolithin C, and ellagic acid.
Claim 5 further limits the method of claim 1 wherein the urolithin is urolithin A, or a combination of Urolithin A with at least one of Urolithin B, Urolithin C or Urolithin D.
As discussed previously, Sanchez teaches administration of compositions comprising urolithin A, urolithin C, and ellagic acid.
Claim 6 further limits the method of claim 1 wherein the urolithin is urolithin A.
As discussed previously, Sanchez teaches administration of compositions comprising urolithin A.
Claim(s) 7 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez in view of Vicinanza.
Claim 7 further limits the method of claim 1 wherein the urolithin is administered in an amount of about 500-2000 mg/day.
As discussed previously, Sanchez essentially teaches a method of preventing recurring cancer by administering a composition comprising ellagic acid and urolithins. Sanchez does not explicitly teach administering urolithin in an amount of 500-2000 mg/day. However, it would have been obvious to do so per the principle of routine optimization because Vicinanza teaches that urolithin administration provides both dose and time dependent effects on cancer proliferation.
As previously indicated in the rejection of claim 3, Vicinanza provides in vitro and ex vivo testing of ellagic acid and urolithin A on cancer proliferation. The results of said testing indicate that both provide dose and time dependent antiproliferative effects (page 5, Fig. 2). Given the evidence provided by Vicinanza, it would have been obvious for a person of ordinary skill in the art to find the optimal dose amounts and intervals required for the effective treatment of a patient and apply such values to the compositions and methods of Sanchez. See MPEP2144.05(II):
Generally, differences in concentration or temperature will not support the patentability of subject matter encompassed by the prior art unless there is evidence indicating such concentration or temperature is critical. "[W]here the general conditions of a claim are disclosed in the prior art, it is not inventive to discover the optimum or workable ranges by routine experimentation." In re Aller, 220 F.2d 454, 456, 105 USPQ 233, 235 (CCPA 1955) (Claimed process which was performed at a temperature between 40°C and 80°C and an acid concentration between 25% and 70% was held to be prima facie obvious over a reference process which differed from the claims only in that the reference process was performed at a temperature of 100°C and an acid concentration of 10%.); see also Peterson, 315 F.3d at 1330, 65 USPQ2d at 1382 ("The normal desire of scientists or artisans to improve upon what is already generally known provides the motivation to determine where in a disclosed set of percentage ranges is the optimum combination of percentages."); In re Hoeschele, 406 F.2d 1403, 160 USPQ 809 (CCPA 1969) (Claimed elastomeric polyurethanes which fell within the broad scope of the references were held to be unpatentable thereover because, among other reasons, there was no evidence of the criticality of the claimed ranges of molecular weight or molar proportions.).
Claim(s) 8-9 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez.
Claim 8 further limits the method of claim 1 wherein the administration of the composition comprising urolithin starts at day 0 of the remission phase or at a later stage.
As discussed previously in the rejection of claim 1, the teachings of Sanchez are also directed towards the prevention of cancer recurrence. The instant claim indicates dosing at day 0 of the remission phase, or a later phase (i.e., any time during remission). Accordingly, the administering of a composition comprising urolithins to a patient in cancer remission would be obvious for the same reasons as indicated in claim 1.
Claim 9 further limits the method of claim 1 wherein the cancer is a solid tumor.
The teachings of Sanchez are directed towards CRC, which would be considered as a type of solid tumor.
Claim(s) 10-11 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez in view of Vicinanza.
Claim 10 further limits the method of claim 1 wherein the preceding cancer therapy is selected from the group consisting of a cancer surgery, a radiation therapy, a chemotherapy, a targeted cancer therapy, an immunotherapy, a hormonal therapy, and a phytochemical based cancer therapy.
As discussed previously, Sanchez obviates a method of claim 1. However, Sanchez does not explicitly teach administration of their urolithin composition to a patient having undergone a preceding cancer therapy chosen from the recited group. However it would be obvious to do so because Vicinanza indicates the administration of pomegranate juice to patients having previously undergone surgery or radiation therapy for prostate cancer (page 1)8. Administration of pomegranate juice to said patients resulted in an increase in doubling time of prostate specific antigens which indicate a slowing of prostate cancer cell growth. Vicinanza attributes this effect to metabolites derived from the pomegranate juice, which include ellagic acid and urolithin A.
Given the teachings of both Sanchez and Vicinanza, it would have been prima facie obvious for a person of ordinary skill in the art to administer the compositions of Sanchez to patients having undergone previous cancer surgery or radiation therapy, because there would be a reasonable expectation that such treatment would be effective in reducing risk of cancer recurrence in such a patient population.
Claim 11 further limits the method of claim 1 wherein the preceding cancer therapy involves use of urolithin.
As discussed previously, Sanchez obviates a method of claim 1. However, Sanchez does not explicitly teach the administration of their compositions after a preceding cancer therapy involving urolithin. However, it would be obvious to do so because Sanchez teaches the treatment of cancer by administering urolithin compositions, while Vicinanza establishes post-therapy administration of pomegranate juice to decrease prostate specific antigen, and there would be a reasonable expectation of success in treating and/or preventing cancer recurrence.
Firstly, the teachings of Sanchez are not only drawn to cancer recurrence, but also provide a suggestion indicating that the administration of ellagic acid and urolithin A is capable of modulating chemoresistance of CSCs, providing potential to increase efficacy of chemotherapeutic drugs such as 5-fluorouracil (page 14)9.
Secondly, the teachings of Vicinanza teach administration of pomegranate juice as a follow-up treatment to cancer surgery and radiotherapy. While Vicinanza does not explicitly teach administration proceeding chemotherapy, it establishes administration of pomegranate juice as a secondary treatment on the basis that is capable of reducing the rate of PSA production, thereby providing antiproliferative effect. Both Vicinanza and Sanchez appear to be unified in at least that both support the anti-proliferative effect of ellagic acid and urolithin A.
Given the teachings of Sanchez and Vicinanza, it would have been prima facie obvious to treat cancer using a combination of chemotherapy and urolithin administration per Sanchez, and provide a secondary follow-up treatment by administering a composition comprising urolithin (such as pomegranate juice), because there would have been a reasonable expectation that such a method would be effective in treating cancer and preventing recurrence of said cancer.
Claim(s) 12-13 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez.
Claim 12 further limits the method of claim 1 wherein the composition is administered orally.
Sanchez indicates that urolithin compositions MPhA and MPhB were administered to cell lines in such a way to mimic conditions wherein ET containing products were orally administered to subjects (page 10)10. Accordingly, oral administration would be obvious to a person of ordinary skill in the art as the teachings of Sanchez are drawn to simulating conditions post-oral intake.
Claim 13 further limits the method of claim 1 wherein the composition is in the form of a pharmaceutical or nutritional composition.
The compositions taught by Sanchez would be considered as pharmaceutical compositions.
Claim(s) 14 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez in view of Vicinanza.
Claim 14 further limits the method of claim 13 wherein the composition comprises an extract selected from the group consisting of pomegranate extract, a tamarind extract or-and a mumijo extract, that provides at least a portion of the at least one urolithin or precursor thereof.
As indicated previously, Sanchez obviates the methods of claims 1 and 14. Sanchez further indicates that pomegranates contain ellagitannins which metabolize to form ellagic acid and urolithins. However Sanchez does not explicitly teach the use of a pomegranate extract. However it would be obvious to administer a composition comprising at least pomegranate extract because
Vicinanza teaches the use of pomegranate juice, which would be considered as an extract of pomegranate.
Claim(s) 15 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez.
Claim 15 further limits the method of claim 1 wherein the compositions comprises the urolithin in isolated form.
Sanchez teaches compositions comprising isolated forms of urolithin (A, B, C, etc.).
Claim(s) 16-17 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez in view of Tummala (Cancer Cell 26, 826–839) and Sun (Public Health Nutrition: 19(8), 1446–1456).
Claim 16 further limits the method of claim 1 wherein the composition furthermore comprises a NAD+ precursor and/or a Vitamin B, wherein the NAD+ precursor is selected from the group consisting Nicotinic Acid, Nicotinamide, Nicotinamide Riboside (NR), Reduced Nicotinamide riboside (NRH), Nicotinamide Mononucleotide (NMN), Nicotinic acid mononucleotide, Nicotinic acid riboside, and mixtures thereof.
As discussed previously, Sanchez obviates a method of claim 1 by teaching the administration of compositions comprising urolithin for reducing the risk of cancer recurrence. Sanchez does not explicitly teach the inclusion of an NAD+ precursor and/or a vitamin B in their compositions. However it would be obvious to include such components because Tummala teaches the use of NAD+ precursors for treating and preventing cancer, while Sun teaches the use of vitamin B for cancer prevention, and there would be a reasonable expectation that a combination of the aforementioned would be successful in reducing risk of recurrence of cancer.
Tummala teaches the prevention of tumorigenesis by administration of nicotinamide riboside (NR). More specifically, the teachings of Tummala are directed towards hepatocellular carcinoma associated with expression of unconventional pre foldin RPB5 interactor (URI) (page 831)11. Furthermore, Tummala indicates that URI expression has an inverse relationship with NAD+ concentrations (page 831)12. Further testing was conducted on mice expressing human URI (hURI), wherein said animals were administered NR by feed. Prolonged NR administration resulted in prevention of tumor development, indicating that restoration of NAD+ pools by NR administration was capable of protecting subjects from hURI-induced DNA damage, preneoplastic lesions, and tumor development. Furthermore, subjects having full blown tumors showed tumor regression after 48 weeks of dosing (page 832)13. Accordingly, it would be obvious for a person of ordinary skill in the art to include NR in the composition of Sanchez because there would be a reasonable expectation of synergistic antiproliferative activity.
Sun teaches the relationship between vitamin B12 intake and colorectal cancer risk. More specifically, Sun provides an assessment of aggregated data from existing studies and provides an analysis of dose response association between B12 intake and relative colorectal cancer risk. Sun indicates that their aggregate data provides a non-linear dose-response curve between vitamin b12 intake and CRC risk as exemplified in the graph below (page 1451, Fig. 2):
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Sun found that vitamin B12 intake was inversely associated with CRC risk, but only significantly when total B12 intake was above 12.85 ug/day (page 1448)14. Accordingly, it would have been obvious for a person of ordinary skill in the art to include vitamin B12 in the composition of Sanchez because there would have been a reasonable expectation of synergistic activity in reducing cancer recurrence.
Given the teachings of Sanchez, Tummala, and Sun, it would have been prima facie obvious for a person of ordinary skill in the art to develop a method of the instant claim by administering a composition comprising urolithins, NAD+ precursors, and vitamin B because there would have been a reasonable expectation that a combination of each of the named components would have been successful in reducing risk of cancer recurrence and proliferation.
Claim 17 further limits the method of claim 1 wherein the composition further comprises nicotinamide riboside and vitamin B12.
As discussed previously in the claim 16 rejection, Sanchez obviates a method of claim 1, while Tummala teaches the use of nicotinamide riboside and Sun teaches the use of vitamin B12. The instant claim is accordingly considered as obvious for the same reasons as iterated in claim 16.
Claim(s) 18 is/are rejected under 35 U.S.C. 103 as being unpatentable over Sanchez.
Claim 18 recites a method of reducing the reducing the risk of the recurrence of cancer or tumor formation in a patient in need thereof in the remission phase after a preceding cancer therapy in said patient, wherein the method comprises the following steps:
Providing a composition comprising a therapeutically effective amount of a Urolithin or a precursor thereof
Administering the composition comprising the therapeutically effective amount of a Urolithin or a precursor thereof to the patient in need thereof, wherein the patient is in the remission phase after a preceding cancer therapy
Monitoring the patient regularly on the potential recurrence of the cancer treated in the preceding cancer therapy
The instant claim is largely identical to the method of claim 1. Essentially, the only difference between the instant claim and claim 1 is the recitation of step c), wherein the patient is monitored for cancer recurrence. While Sanchez does not explicitly teach the monitoring of a patient, such actions would be obvious and commonsense in the field of medicine and pharmaceuticals. A person of ordinary skill in the art would naturally choose to monitor a patient undergoing treatment, especially when the target disease of the said treatment is recurrence of cancer previously treated, and the patient is at risk of relapsing.
Conclusion
Claims 1-18 are rejected.
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/ERIC TRAN/ Examiner, Art Unit 1629
/JEFFREY S LUNDGREN/ Supervisory Patent Examiner, Art Unit 1629
1 “These mixtures reduce phenotypic and molecular features in two models of colon CSCs: Caco-2 cells and primary tumour cells from a patient with CRC. The mixture containing mostly Uro-A (85% Uro-A, 10% Uro-C, 5% EA) was most effective at inhibiting the number and size of colonospheres and aldehyde dehydrogenase activity (ALDH, a marker of chemoresistance) whereas the mixture containing less Uro-A but IsoUro-A and Uro-B (30% Uro-A, 50% IsoUro-A,10% Uro-B, 5% Uro-C, 5% EA) had some effects on the number and size of colonospheres but not on ALDH. These data support a role for polyphenols metabolites in the control of colon cancer chemoresistance and relapse and encourage the research on the effects of polyphenols against CSCs.”
2 “In this exploratory study we present the first evidence that mixed ETs gut microbiota-derived metabolites, urolithins and EA, have an inhibitory effect on specific phenotypic (colonosphere formation and growth) and molecular features (ALDH activity) attributed to colon CSCs.”
3 “In the specific case of colorectal cancer (CRC), a wide range of polyphenolic compounds and some of their colonic derived metabolites have been shown to be effective against well-established colon cancer cell lines and animal models of CRC (as reviewed by Núnez-Sanchez et al., 2015). It is thus possible that dietary polyphenols and their metabolites may also target colorectal CSCs and therefore help prevent colon cancer metastasis and (or) recurrence.”
4 “We now report that mixed ET-derived metabolites at mM concentrations that can be attained in the colon following oral intake of ET-containing products and in proportions that repro duce the metabolic profile of compounds found in the colon of humans, also exert an effect against CSCs. These experimental conditions resemble more closely those that can be found in vivo and reinforce the relevance of these results.”
5 “Ellagitannins (ETs) from pomegranate juice (PJ) are bioactive polyphenols with chemopreventive potential against prostate cancer (PCa). ETs are not absorbed intact but are partially hydrolyzed in the gut to ellagic acid (EA). Colonic microflora can convert EA to urolithin A (UA), and EA and UA enter the circulation after PJ consumption.”
6 “The sensitivity of cell growth inhibition in the presence of increasing concentrations of EA (from15to60𝜇mol/L) and UA (from 15 to 90𝜇mol/L) was examined at 24, 48, 72, 96, and 120 hours in DU-145 and PC 3, two androgen-independent PCa cell lines. The treatments with EA and UA decreased cell growth in both”
7 “Accumulating experimental evidence has demonstrated that PE inhibits tumor angiogenesis[12], delays the transition from androgen-dependent to androgen-independent phenotype, and induces apoptosis through a nuclear factor-kB dependent mechanism in vitro and in tumor tissue excised from castrated severe combined immunodeficiency (SCID) mice with Los Angeles Prostate Cancer-4 (LAPC-4) human PCa xenografts [13].”
8 “Under these circumstances, rising PSA represents tumor growth, and men with shorter doubling times of PSA value are presumed to have more rapidly growing tumors[4, 5]. A phase II study examining the effects of pomegranate juice (PJ) in men with rising PSA following surgery or radiation for PCa demonstrated that consumption of 8 ounces of PJ significantly increased the PSA doubling time, from 15 to 54 months, suggesting an inhibitory action of PJ metabolites on PCa cell growth [6].”
9 “Since high ALDH activity is associated with the chemo resistance of colon cancer cells (Dylla et al., 2008; Nautiyal et al., 2011) our results and that of others (Kakarala et al., 2010; Li et al., 2010; Mineva et al., 2013) support a potential modulatory effect of polyphenols and derived metabolites on the chemoresistance of cancer cells. It is worth mentioning that the increased resistance of CSCs to commonly used chemotherapeutic drugs is correlated with a higher drug-efflux capacity, mediated by a higher expression of ABC transporter proteins in these cells (Zhou et al., 2001). In this regard, Uro-A has been reported to be substrate of ABCG2/BCRP (Gonzalez-Sarrías et al., 2013) and to potentiate the anticancer effects of 5-fluorouracil (5-FU) against human colon cancer cells (Gonzalez-Sarrías et al., 2015c). Further investigations are needed to determine the efficacy of combined urolithins and 5-FU against the growth and chemoresistance of CSCs.”
10 “Cells were treated with each of two mixtures of metabolites (MPh) that were prepared to mimic qualitatively and quantitatively the profile of compounds detected in the human colon and faeces following oral intake of ET-containing products”
11 “In sequential immunoprecipitation experiments, using 1-week-old liver ex tracts, free hURI molecules were revealed by WB after complete depletion of PP1g (Figures S4B and S4C), and vice versa (data not shown). When 3-week-old mice were supplied a rapamycin-containing diet, progression to preneoplastic abnormalities continued, if not further pronounced (data not shown). Thus, although a fraction of hURI binds PP1g, hURI apparently has a PP1g-independent role in DNA damage and liver tumorigenesis. Additionally, no differences in reactive oxygen species (ROS) were observed in 1- and 8-week-old livers (Figures S4D and S4E), suggesting that DNA damage is ROS-independent.”
12 “Furthermore, NAD+ concentrations inversely correlated with URI levels in four human HCC cell lines (Huh-7, HepG2, SNU-398, and SNU-449). While URI depletion significantly increased NAD+ levels, URI overexpression reduced NAD+ values (Figure S4P). URI overexpression in SNU-449 cells, which had high NAD+ values and low endogenous URI levels, increased their growth, whereas URI depletion in Huh-7 and HepG2 cells displaying high endogenous URI, significantly reduced their growth (Figure S4Q). URI-regulating NAD+ levels may therefore be relevant for human liver tumorigenesis.”
13 “Prolonged NR treatment pre vented tumor development and reduced ALT levels (Figures 5E–5G). Similarly liver tumors were prevented in 30-week-old homozygous mutants with higher URI levels (Figure S5E). Thus, restoring NAD+ pools protects from hURI-induced DNA damage, preneoplastic lesions, and tumor development. Surprisingly, 12-week-old homozygous mutants with full blown tumors then on 48 weeks of NR regimen showed significant tumor regression”
14 “In the non-linear dose–response relationship, there was a slight trend towards a reduction in CRC risk when vitamin B12 intake was less than 12·85μg/d, although the reduction was not statistically significant, and a significantly reduced risk was observed when vitamin B12 intake was more than 12·85μg/d.”
Prosecution Insights