Introduction:

Molecular diagnostics using RT-PCR has now emerged as the new diagnostic method for clinicians with the dawn of SARS-CoV-2. In India, the popularity and awareness of RT-PCR and particularly the increased availability of testing machines across hospitals has now opened up possibilities of diagnostic tests with RT-PCR. In view of the cytokine storm which is the significant reason for morbidity and mortality of COVID-19 patients, we proposed to test the usefulness of a multiplex RT-PCR test kit that simultaneously measured inflammatory markers namely, IL-6, TNF– α and IL-10 (IFM) all in one tube. The study included a group of patients who were equally allotted to two treatment arms one of which received standard of care along with a food supplement capsule as a natural anti-inflammatory (RECOVEREEZ FORTE™), and the other group received standard of care that included oral Prednisolone tablets in tapered dosage. RECOVEREEZ FORTE™ consists of potent biomolecules from cardamom extract. A natural product with substantial anti-inflammatory action when consumed early at the onset of symptoms is hereby proven by comparing gene expression profile of inflammatory markers with routinely tested inflammatory parameters such as serum IL-6, CRP and LDH. In addition to predicting worse disease outcomes beforehand, RT-PCR assay tests provides an opportunity for identifying therapeutic window aiding in practicing effective treatment strategy for COVID-19. RT-PCR analysis of IFM together being used in a single multiplex kit is being first reported and such a test as a prognosticator for disease progression does seem promising and worthy of clinicians adopting novel testing modalities in clinical practice.

Aim of the study:

To study the anti-inflammatory response of RECOVEREEZ FORTE™ using RT-PCR based multiplex gene profiling of inflammatory markers in disease prognosis and show its predictability of worsening outcomes and its role in identifying therapeutic window for RECOVEREEZ FORTE™. To show the potentiality of RECOVEREEZ FORTE™ in reducing COVID-19 symptoms and SARS-CoV-2 RT-PCR Ct values of the treatment group. Materials and methods This was a short study of a 10-day period where the end point was the negativity of COVID-19 on RT-PCR test or the decline in cycle threshold (Ct) values of the test performed on day 10 when compared to day 0. During the course of the study, patients were given standard treatment and an oral dose of 500 mg of RECOVEREEZ FORTE™ thrice daily, or standard of care and oral Prednisolone in tapered dosage (control group). All essential interventions were
included in the standard of care as decided by the attending physician. The RT-PCR results of inflammatory markers were compared to routinely tested parameters such as IL-6, CRP and LDH. Time to clinical improvement was in terms of SARSCoV-2 RT-PCR test negativity or recovery of COVID-19 symptoms. We enrolled 64 patients, of which 32 were allocated to RECOVEREEZ FORTE ™ group and 32 to the control group.

Results:

The RT-PCR analysis of elevated IFM on day 0 corresponded to above normal protein levels of routinely tested parameters such as serum IL-6, CRP and LDH on day 5. Similarly, elevated IFM on day 5 corresponded to above normal protein levels on day 10. Such an association was equally prevalent in both the control group and RECOVEREEZ FORTE™ group, stipulating that RECOVEREEZ FORTE™ may be used as an alternative to steroids. The obtained results indicate that the RT-PCR assessment predicts worse outcomes 5 days earlier. But, the RT-PCR analysis of elevated IFM on day 0 did not correspond to the above normal protein levels of other parameters on day 10. Hence, indicating that the IFM RT-PCR test cannot predict worse outcomes 10 days earlier. Also, RECOVEREEZ FORTE™ when consumed for a period of 10 days normalized LDH values, compared to the control group. Moreover, IFM RT-PCR test identified a 5-day therapeutic window for RECOVEREEZ FORTE™ against inflammation experienced by patients. In addition to the above findings, the authors also observed that majority of the patients belonging to the treatment group showed recovery from symptoms such as fever, cough, sore throat and breathlessness compared to control group by day 5. 12 (37.5%) out of 32 patients and 6 (18.75%) out of 32 patients belonging to the treatment group and control group, respectively, became SARS-CoV-2 negative by day 5, indicating a probable anti-viral action of RECOVEREEZ FORTE™ against SARS-CoV-2. Conclusion The IFM RT-PCR test possess 5-day early prediction ability and lacks 10-day prediction ability. Treatment with RECOVEREEZ FORTE™ indicates good anti-inflammatory action which is equivalent to steroids. Intake of RECOVEREEZ FORTE ™ for a period of 5 days depicts persistent anti-inflammatory action, recovery of COVID-19 symptoms and a probable anti-viral action. Moreover, an effective normalization of LDH may be rendered by RECOVEREEZ FORTE™ when consumed for a period of 10 days.

Keywords: COVID-19 · Inflammation · Prognosis · IL-6 · CRP · LDH · Cardamom

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) provokes demanding immune and inflammatory events. Despite numerous reports on the use and testing of several potent therapeutic options against coronavirus disease 2019 (COVID-19), satisfactory treatment has not so far been determined. RECOVEREEZ FORTE TM, consisting of a standardized cardamom extract is a natural product with substantial indications of immunomodulatory and anti-inflammatory actions along with inhibitory capacity averse to viral targets. In this context, we speculated that RECOVEREEZ FORTE TM may ameliorate adverse effects in COVID-19 patients. Accordingly, in a multicenter prospective, open label, randomized trial, adult COVID-19 patients having mild to moderate symptoms were treated with RECOVEREEZ FORTE TM as an adjunct therapy. Patients were assigned to obtain standard of care along with a three times per day oral dose of 500 mg of RECOVEREEZ FORTE TM for ten days, or standard of care alone. Standard of care comprised all essential interventions, as per the discretion of the attending physician. Time to clinical improvement in terms of biochemical parameters such as IL6, CRP, DDIMER, LDH and an RTPCR COVID-19 test negativity in the treated patients was considered to be the primary end point. We enrolled 60 patients; of which 30 were allocated to RECOVEREEZ FORTE TM and 30 to the control group. The duration of COVID-19 positivity post-intervention was significantly shorter in RECOVEREEZ FORTE TM treated group than in the control group (p<0.001). RECOVEREEZ FORTE TM group also showed significant decrease in the levels of IL6, LDH, D-dimer along with increased lymphocytes confirming its immunomodulatory and anti-inflammatory action. We conclude that RECOVEREEZ FORTE TM can reduce the impact of COVID-19 manifestations and could serve as an alternative to oral steroids. We also suggest that RECOVEREEZ FORTE TM may aid faster recovery mediated by its immunomodulatory and anti-inflammatory action in diseases other than COVID-19. The trial was registered in Clinical Trials Registry India (CTRI – CTRI/2021/04/033143) in compliance with the International Council on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use – Good Clinical Practice (ICH–GCP) guidelines.

KEYWORDS: RECOVEREEZ FORTE TM, COVID-19, Anti-inflammatory action, Immunomodulatory action,
Anti-viral Activity, Cardamom extracts