Stress test and treatment of chronic kidney disease
By administering a transient oxidative stress inducer to stimulate antioxidant protein expression and measuring their levels, the method quantitatively assesses and addresses deficiencies in antioxidant capacity, guiding personalized treatment for chronic kidney disease.
Patent Information
- Application Number
- JP2025112819
- Authority / Receiving Office
- JP · JP
- Patent Type
- Applications
- Current Assignee / Owner
- Priority Date
- 2020-03-23
- Filing Date
- 2025-07-03
- Publication Date
- 2025-09-25
AI Technical Summary
Current methods lack the ability to predictively assess antioxidant defense in humans, particularly in patients with chronic kidney disease, necessitating a need for quantitative measurement and therapeutic strategies based on antioxidant response.
Administering a transient oxidative stress inducer, such as protoporphyrin, to patients to stimulate the expression of antioxidant proteins like HO-1, ferritin, or NQO1, followed by measuring their levels to determine the patient's antioxidant reserve, and administering a treatment if the levels exceed a predefined threshold.
Provides a quantitative method to assess and address deficiencies in antioxidant capacity, guiding personalized treatment strategies for chronic kidney disease by using potent antioxidants like tetrahydrocurcumin or Nrf2 activators.
Smart Images
Figure 2025138830000001_ABST
Abstract
Description
[Technical Field]
[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 993,446, filed March 23, 2020, which is incorporated herein by reference. [Background technology]
[0002] Oxidative stress is a hallmark and mediator of chronic kidney disease (CKD). A decline in antioxidant defense capacity is thought to be part of the cause. However, there are currently no methods for predictively assessing antioxidant defense in humans. Therefore, there is a need for quantitative methods for measuring antioxidant defense in patients and for therapeutic strategies that take into account the level of a patient's antioxidant response. Summary of the Invention [Means for solving the problem]
[0003] (Summary of the Invention) The present invention relates to a method for treating a patient, comprising: (a) administering to the patient a compound that is a transient oxidative stress inducer; (b) measuring the patient's response to the compound, wherein the response comprises an elevated level of expression of one or more antioxidant proteins; and (c) administering a treatment to the patient if the expression level of the one or more antioxidant proteins exceeds a predefined level. In one embodiment, the patient may be suffering from chronic kidney disease. The compound may be a protoporphyrin, such as tin protoporphyrin, and the antioxidant proteins include one or more of HO-1, ferritin, p21, or NQO1. The treatment may comprise administering an antioxidant, such as tetrahydrocurcumin.
[0004] In one embodiment, the method is carried out such that the expression level of one or more antioxidant proteins is measured before step (a), and the expression level of one or more antioxidant proteins measured in step (b) is compared to the expression level measured before step (a).
[0005] In another aspect, the present invention relates to a method comprising: (a) administering a compound to a patient, whereby the compound stimulates the production of one or more antioxidant proteins in the patient; (b) obtaining one or more body fluid samples from the patient; and (c) measuring the level of the one or more antioxidant proteins produced in the patient. The compound may be a transient oxidative stress inducer, including a protoporphyrin or mesoporphyrin, such as tin protoporphyrin. The antioxidant comprises one or more of HO-1, ferritin, p21, or NQO1. In another aspect, the production level of the one or more antioxidant proteins is measured before step (a), and the expression level of the one or more antioxidant proteins measured in step (c) is compared to the expression level measured before step (a). The measured antioxidant protein level provides a quantitative indicator of the patient's antioxidant reserve. [Brief explanation of the drawings]
[0006] [Figure 1] 1 shows that plasma HO-1 levels increase in a dose-dependent manner after SnPP injection in healthy volunteers. [Figure 2] We show that plasma HO-1 levels are elevated after SnPP injection in participants with stage 3 CKD (15-29 ml / min / 1.73 m2) and stage 4 CKD (30-59 ml / min / 1.73 m2). [Figure 3] 1 shows that plasma ferritin levels increase in response to SnPP injection. [Figure 4] 1 shows baseline NQO1 levels in healthy volunteers and participants with CKD. [Figure 5] Plasma and urinary NQO1 responses to 90 mg SnPP infusion are shown. [Figure 6] Plasma p21 concentrations at baseline and after SnPP injection are shown. [Figure 7] Baseline and peak plasma values of the proteins tested are shown, indicating the degree of response to the highest tested dose of SnPP (90 mg). DETAILED DESCRIPTION OF THE INVENTION
[0007] The inventors have determined that SnPPs elevate various stress proteins in humans, thereby serving as a pharmaceutical "stress test" for measuring human genetic responsiveness and, therefore, antioxidant reserve. Stress testing can be part of a method for treating chronic kidney disease by administering potent antioxidants to patients whose tests indicate a deficiency in antioxidant reserve. The test can also be used to independently assess a patient's antioxidant reserve level to further guide their treatment.
[0008] In one aspect, the present invention includes a method of treating a patient comprising the steps of: (a) administering to the patient a compound that is a transient oxidative stress inducer; (b) measuring the patient's response to the compound, the response comprising an elevated level of expression of one or more antioxidant proteins; and (c) administering a treatment to the patient if the expression level of one or more antioxidant proteins is above a predefined level.
[0009] The compound is preferably a protoporphyrin or mesoporphyrin, such as tin protoporphyrin. The compound may also be zinc, tin, or cobalt protoporphyrin or mesoporphyrin. The compound must be capable of inducing the production of stress proteins in humans in a dose-dependent manner. This allows for quantitative measurement of the patient's antioxidant reserve capacity. Antioxidant proteins include one or more of HO-1, ferritin, p21, or NQO1. In one embodiment, the present invention measures the patient's baseline levels of these proteins before administering the compound. This allows for comparison of the increased levels of proteins induced by administering the compound.
[0010] Certain methods disclosed herein are useful for treating conditions associated with chronic kidney disease. When patients being treated for chronic kidney disease lack antioxidant reserve, they may benefit from the administration of potent antioxidants, such as tetrahydrocurcumin (19, 20), or Nrf2 activators, such as bardoxolone methyl.
[0011] Another aspect of the present invention is a method that allows measuring the antioxidant capacity of a patient in a dose-dependent manner, the method comprising the following steps: (a) administering a compound to a patient, whereby the compound stimulates the production of one or more antioxidant proteins in the patient; (b) obtaining one or more bodily fluid samples from the patient; and (c) measuring the level of one or more antioxidant proteins produced in the patient.
[0012] The compound is preferably a transient oxidative stress inducer, such as tin protoporphyrin or other protoporphyrins or mesoporphyrins. Administration of the compound induces stress proteins, such as one or more of HO-1, ferritin, p21, or NQO1. The induced levels of these proteins provide a quantitative indicator of the patient's antioxidant reserve capacity. Those patients with a capacity lower than a predefined value are candidates for potent antioxidant therapy.
[0013] As shown in Figure 1, plasma HO-1 levels increased dose-dependently after SnPP injection in healthy volunteers. Plasma HO-1 levels were measured at baseline and on days 1-4 after injection of 9, 27, or 90 mg of SnPP. A time- and dose-dependent increase was observed (mean values / 95% confidence intervals shown). Significant increases in HO-1 levels over time were observed in each subject group (ANOVA / repeated measures; all p values <0.001 as determined by within-group analysis).
[0014] As shown in Figure 2, plasma HO-1 levels increased after SnPP injection in participants with stage 3 CKD (15-29 ml / min / 1.73 m²) and stage 4 CKD (30-59 ml / min / 1.73 m²). Plasma HO-1 levels were measured at baseline and on days 1-4 after injection of 27 mg or 90 mg of SnPP. A time- and dose-dependent increase in HO-1 was observed in both the CKD3 and CKD4 groups. P < 0.001 for all between-group comparisons; ANOVA, repeated measures. Mean values / 95% confidence intervals are shown.
[0015] As shown in Figure 3, plasma ferritin levels increased in response to SnPP injection. Plasma ferritin levels were measured at baseline and 24+48 hours after 90 mg SnPP injection in healthy volunteers and CKD3 / CKD4 participants. CKD participants had significantly higher baseline ferritin levels than healthy volunteers (p<0.005 for between-group comparisons). All three groups showed a significant increase in plasma ferritin after SnPP injection (p-values by ANOVA, repeated measures, within-group analysis). The increase relative to baseline values was similar in the CKD and healthy volunteer groups. Means and 95% confidence intervals are shown.
[0016] Baseline NQO1 levels in healthy volunteers and CKD participants are shown in Figure 4. The combined CKD cohort and healthy volunteers had virtually identical plasma NQO1 concentrations. However, CKD patients had significantly elevated urinary NQO1 / creatinine levels (p<0.01; unpaired t-test). When urinary NQO1 / creatinine values were transformed to log base 10, individual values showed a strong inverse correlation with each subject's corresponding baseline eGFR (r, -0.85).
[0017] The plasma and urinary NQO1 responses to 90 mg SnPP infusion are shown in Figure 5. Healthy volunteers and CKD participants showed acute and nearly identical increases in plasma NQO1 after 90 mg SnPP injection. Furthermore, healthy volunteers and combined CKD participants showed significant and progressive increases in urinary NQO1 / creatinine levels over the 72-hour observation period. Although healthy volunteers had lower baseline urinary NQO1 levels than CKD participants, the slope of the SnPP-induced increase in urinary NQO1 over time was similar in the healthy volunteer and CKD groups. Values are means / 95% confidence intervals; urine values were log-base 10 transformed; ANOVA with repeated measures, within-group comparisons.
[0018] Plasma p21 concentrations at baseline and after SnPP injection are shown in Figure 6. There was significant variability in p21 values between participants, as reflected by the large standard deviation (SD; numbers in parentheses shown at the bottom of the graph). (HV = healthy volunteers, BL = baseline). However, despite this variability, both healthy and CKD participants showed elevated plasma p21 levels at 12 and 24 hours after SnPP injection (CKD participants combined, p < 0.01; healthy volunteers, p < 0.01; ANOVA, repeated measures, between-group comparison). The fold increase relative to baseline values was significantly higher in the healthy volunteer group than in the CKD group (mean 2.65 vs. 0.51, respectively, p < 0.001; see text).
[0019] [Table 1]
[0020] Legend for Table 1. Demographic and baseline clinical data for the three study cohorts. Means and standard deviations (numbers in parentheses) are shown. CKD3, 15–29 ml / min / 1.73 m 2 CKD4, 30-59 ml / min / 1.73 m 2 For the three classes of medications, the percentage of participants taking them within each group is shown. BP, blood pressure; BUN, blood urea nitrogen.
[0021] [Table 2]
[0022] Legend for Table 2. eGFR is ml / min / 1.73m 2 CKD3 = eGFR between 30 and 59 ml / min / m 2 CKD4 = eGFR 5-29 ml / min / m 2 Values are presented as mean ± 1 SD. There was no significant change in eGFR from baseline in response to the highest tested dose of tin protoporphyrin (90 mg).
[0023] [Table 3]
[0024] Legend for Table 3. Urinary biomarker values factored by urinary creatinine (Cr). HV = healthy volunteers, CKD3 = CKD stage 3 (eGFR 30-59 ml / min / 1.73 m 2 ), CKD4 = CKD stage 4 (eGFR 15-29 ml / min / 1.73 m 2 ). eGFR was assessed at baseline and on days 1-4 after SnPP injection. No significant changes were observed over time for any analyte (see p-values). KIM-1 = kidney injury molecule 1, NGAL = neutrophil gelatinase-associated lipocalin, NAG = n-acetylglucosaminidase. All values = mean ± 1 SD. p-values are shown for changes over time.
[0025] Figure 7 shows baseline and peak plasma values of the tested proteins, indicating the degree of response to the highest tested dose of SnPP (90 mg). Baseline and maximum (peak) plasma antioxidant protein concentrations produced using the highest tested dose of SnPP (90 mg) are shown. CKD = CKD3 and CKD4 groups combined. Data indicate the ability of SnPP to upregulate HO-1, ferritin, NQO1, and p21 gene / protein expression. Values shown are means and 95% confidence intervals. See figures for changes in values over time and statistics from repeated measures ANOVA. All peak values were significantly higher than baseline values, but the CKD group showed a significantly blunted p21 response compared to the healthy volunteer group (see text). The timing of peak values was 4 days, 12 hours, 12 hours, and 4 hours for HO-1, ferritin, p21, and NQO1, respectively.
[0026] Other embodiments and applications of the present invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. All references cited herein, including all U.S. and foreign patents and patent applications, are specifically and entirely incorporated herein by reference. It is intended that the specification and examples be considered as exemplary only, with the true scope and spirit of the invention being indicated by the following claims.
Claims
1. 1. A method for treating a patient, comprising: (a) administering to the patient a compound that is a transient oxidative stress inducer; (b) measuring the patient's response to the compound, the response comprising an elevated level of expression of one or more antioxidant proteins; and (c) administering a treatment to the patient if the expression level of one or more antioxidant proteins is above a predefined level. A method comprising:
2. 2. The method of claim 1, wherein the antioxidant protein comprises one or more of HO-1, ferritin, p21, or NQO1.
3. 3. The method of claim 1 or 2, wherein the expression levels of one or more antioxidant proteins are measured before step (a), and the expression levels of one or more antioxidant proteins measured in step (b) are compared with the expression levels measured before step (a).
4. The method according to any one of claims 1 to 3, wherein the patient is suffering from chronic kidney disease.
5. The method according to any one of claims 1 to 4, wherein the compound is protoporphyrin or mesoporphyrin.
6. The method of any one of claims 1 to 5, wherein the compound is a metalloprotoporphyrin.
7. The method according to any one of claims 1 to 6, wherein the metalloprotoporphyrin is tin, cobalt, or zinc protoporphyrin.
8. The method of any one of claims 1 to 7, wherein the dose of the compound is 9 mg or more.
9. 9. The method of any one of claims 1 to 8, wherein the treatment is a strong antioxidant.
10. 10. The method of any one of claims 1 to 9, wherein the treatment comprises administering tetrahydrocurcumin.
11. (a) administering a compound to a patient, whereby the compound stimulates the production of one or more antioxidant proteins in the patient; (b) obtaining one or more bodily fluid samples from the patient; and (c) measuring the level of one or more antioxidant proteins produced in the patient; A method comprising:
12. 12. The method of claim 11, wherein the compound is a transient oxidative stress inducer.
13. 13. The method of claim 11 or 12, wherein the compound is protoporphyrin or mesoporphyrin.
14. The method according to any one of claims 11 to 13, wherein the compound is a metalloprotoporphyrin.
15. The method according to any one of claims 11 to 14, wherein the metalloprotoporphyrin is tin, cobalt, or zinc protoporphyrin.
16. The method of any one of claims 11 to 15, wherein the antioxidant protein comprises one or more of HO-1, ferritin, p21, or NQO1.
17. 17. The method of any one of claims 11 to 16, wherein the production level of one or more antioxidant proteins is measured before step (a), and the expression level of one or more antioxidant proteins measured in step (c) is compared with the expression level measured before step (a).
18. The method of any of claims 11 to 17, wherein the measured antioxidant protein levels provide a quantitative indication of the patient's antioxidant reserve.