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Citation: Herscu, P.; Talele, G.;
Vaidya, S.; Shah, R. Safety and
Evaluation of the Immune Response
of Coronavirus Nosode
(BiosimCovex) in Healthy Volunteers:
A Preliminary Study Extending the
Homeopathic Pathogenetic Trial.
Medicines 2023,10, 8. https://
doi.org/10.3390/medicines10010008
Academic Editor: Hiroshi Sakagami
Received: 12 December 2022
Revised: 21 December 2022
Accepted: 26 December 2022
Published: 30 December 2022
Copyright: © 2022 by the authors.
Licensee MDPI, Basel, Switzerland.
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).
medicines
Article
Safety and Evaluation of the Immune Response of Coronavirus
Nosode (BiosimCovex) in Healthy Volunteers: A Preliminary
Study Extending the Homeopathic Pathogenetic Trial
Paul Herscu 1, Gitanjali Talele 2, Shashikant Vaidya 3and Rajesh Shah 2 ,*
1Herscu Laboratory, Research Division, 356 Middle Street, Amherst, MA 01002, USA
2Life Force Foundation Trust, 411 Krushal Commercial Complex, Chembur,
Mumbai 400089, Maharashtra, India
3
Assistant Director and HOD Microbiology Department, Haffkine Institute for Training Research and Testing,
Acharya Dhonde Marg, Parel Village, Parmanand Wadi, Parel, Mumbai 400012, Maharashtra, India
*Correspondence: sanjivak@gmail.com
Abstract: Objectives
: Regulatory clinical Phase I studies are aimed at establishing the human safety
of an active pharmaceutical agent to be later marketed as a drug. Since homeopathic medicines are
prepared by a potentizing method using alcohol, past a certain dilution, their toxicity/infectivity is
assumed to be unlikely. We aimed to develop a bridge study between homeopathic pathogenetic
trials and clinical trials. The primary purpose was to evaluate the safety of a nosode, developed from
clinical samples of a COVID-19 patient. The secondary objectives were to explore whether a nosode
developed for a specific clinical purpose, such as use during an epidemic, may elicit laboratory signals
worthy of further exploration.
Methods
: An open-label study was designed to evaluate the safety
and immune response of the Coronavirus nosode BiosimCovex, given orally on three consecutive
days to ten healthy volunteers. Clinical examinations, laboratory safety and immune parameters were
established. Interferon–gamma, Interleukin-6, and CD 4 were measured. (CTRI registration number:
CTRI/2020/05/025496).
Results
: No serious/fatal adverse events were reported. Laboratory tests
to measure safety were unchanged. Three subjects showed elevated Interleukin-6 (IL-6) on day
17 in comparison to the baseline, and ten subjects showed elevated IL-6 on day 34. A significant
difference between IL-6 observations, calculated by repeated measures ANOVA, was found to be
highly significant. On day 60, the IL-6 values of nine subjects were found to return to normal.
Corresponding CD4 cell elevation was observed on day 60, when compared to day 34.
Conclusions
:
HPT may potentially extend into physiological changes with regards to immune response and should
encourage future studies.
Keywords:
coronavirus; SARS-CoV-2; homeopathic pathogenetic trials (HPT); COVID-19; Biosim-
Covex; nosode; potentization; homeopathy; safety; immune response; phase-1 study
1. Introduction
Clinical Phase I trials for the regulatory approval of drugs are generally small and are
conducted to primarily examine the safety of new medicines in healthy subjects before pro-
ceeding further into clinical trials that modify disease. The treatment of illnesses caused by
SARS-CoV-2 is always potentially limited by the toxicity of medications [
1
]. The continued
need for supportive prophylaxis measures, immunotherapeutics and immunomodulators
which may contribute to controlling pandemics; therefore, we considered the development
of potentized preparations (nosodes) [2].
In the homeopathic system of medicine, the drugs are potentized via dilution and
succussion, rendering the source material open to potential nanoparticles [
3
]. It is assumed
that there is little or no toxicological risk remaining past a certain potency when consumed
by human subjects. For example, the potentization process leading to a 30C potency, has
Medicines 2023,10, 8. https://doi.org/10.3390/medicines10010008 https://www.mdpi.com/journal/medicines
Medicines 2023,10, 8 2 of 12
led to an extreme dilution, making the concentration difficult to measure In addition, the
process kills any organism in the original material (bacteria or virus, in the case of nosodes)
due to the continued and repeated exposure of the organism to approximately 90% alcohol,
used as a medium for dilution at every step. As a consequence, human trials for safety
have typically not been a practice for new or old homeopathic drug discovery. Instead,
the model of the Homeopathic Pathogenetic Trial (HPT/drug proving) was developed
to understand the primary effects of the drug substance by recording symptoms during
the trial, which help determine the therapeutic indications, based on the fundamental
homeopathic principle of the law of Similars. Such primary effects are recorded in the
source books, such as Materia Medica Pura (by Samuel Hahnemann [
3
]), Encyclopedia
of Materia Medica (by TF Allen [
4
]) and Hering Guiding Symptoms (by Constantine
Hering [5]).
There are nearly 1500 drug substances recorded in various homeopathic pharma-
copeias (Indian, British, German, and American Pharmacopeia) [
6
,
7
] that are fully or partly
‘proved’ in human trials and have been used by practitioners the world over for the past two
centuries. They are made available across homeopathic pharmacies, as over-the-counter
products, or as dietary supplements, depending on national regulatory requirements. Po-
tentized homeopathic drugs that have been in use for over one hundred years were not
required to undergo any toxicity studies in animals or safety studies in humans.
As an example, ‘The Drug and Cosmetic Act’ (India) states that, to be included in the
pharmacopeia, the drug must be along-used homeopathic drug whose therapeutic efficacy
is recorded in data and established by long clinical use [
8
]. The Act does not ask for animal
or human toxicity studies for homeopathic medicines.
Controversy remains surrounding nosode use as a potential disease prophylactic
during epidemics. Loeb et al. [
9
] showed that homeopathic ‘vaccines’ do not elicit antibody
responses against diphtheria, pertussis, tetanus, and MMR. If nosodes provide some
level of protection during epidemics, they may do so by a mechanism different to that of
conventional vaccines.
The aim of this study is to bridge the homeopathic pathogenetic trial [
10
] technique
with the safety testing commonly found in Phase I drug trials; this study tests the new
medicine BiosimCovex (Coronavirus nosode/CVN01). Secondly, since this medicine is
developed for its potential use as a protective prophylaxis in the current COVID-19 pan-
demic, we examined whether a homeopathically-prepared medicine may elicit certain
blood value changes related to immune response, which may signal potential therapeutic
use. Specifically, with a need for generating scientific data, the present study does not only
investigate IgG/IgM levels against the SARS-CoV-2 Spike Protein, but also includes other
immune parameters that may potentially impact disease presentation. We included the
study of relevant immunomodulatory effects. IL-6 promotes the specific differentiation
of naive CD4 T cells, thus performing an important function in linking the innate to the
acquired immune response [11].
Objective
To evaluate the safety of BiosimCovex in healthy volunteers, this study evaluated
potential immune response signals and recorded subjective symptoms elicited in the trial,
historically and commonly used in homeopathic pathogenetic trials.
2. Materials and Methods
2.1. BiosimCovex (Coronavirus Nosode)
A Coronavirus nosode, initially coded as CVN01 and subsequently labeled as Biosim-
Covex, was prepared from a clinical sample [
12
] of an oropharyngeal swab of a patient;
its use had been approved by the institutional Ethics and Biosafety Committee having
confirmed, by RT-PCR and genome sequencing, the infection of SARS-CoV-2 at Haffkine
Institute, Mumbai. This followed the nosode-making guidelines, as per the Homeopathy
Pharmacopoeia of India, and the Standard Operating Procedure, approved by the Life Force
Medicines 2023,10, 8 3 of 12
Foundation Trust’s Scientific Advisory Body [
13
,
14
]. This nosode is prepared from a clinical
sample, as mentioned in the N-IV group in the HPI [
13
]. Almost all the nosodes (such
as Psorinum, Medorrhinum, Tuberculinum Carcinosin, Syphilinum) in the homeopathic
literature and available on the market have been made from the clinical samples [
15
]. The
ethical and biosafety approvals for the preparation of the nosode were obtained from the
applicable institutional committees. The samples were handled, and the potencies were
prepared in a Biosafety Level-2 (BSL-2) containment lab with BSL-3 practices. Standard
precautions, as per biosafety requirements, were followed during the preparation process.
The clinical sample selected to prepare the nosode had the lower cycle threshold (CT)
and was identified/characterized by gene sequencing. The CT value was approximately 17
and corresponded to 4–9 log10, amounting to the count of more than 20 billion. Based on
the CT value of the sample, the corresponding viral copies, as per the published literature,
were sufficient to start the homeopathy potencies [
16
]. Potencies 1C to 4C were prepared
by using Water for Injection (WFI) as a vehicle. In total, 0.03 mL of OSN was taken in
a suitable glass bottle, allowing one-third space for succussion and the 2.97 mL of WFI
that was added to make a 3 mL volume. The bottle was given 10 strokes with the help
of a hand-potentizer in a bio-safety cabinet to arrive at 1C potency. As per the method of
preparation described in the HPI, the 1:99 ratio was maintained for further potencies. The
nosode, in its final preparation, was delivered on a dry size 30 globule, without alcohol
present. Finally, for safety, RT-PCR confirmed that no Coronavirus was detected beyond
the 3C potency. To document that usable 30C potency is also devoid of any viral material,
above 3C potencies were also tested by using RT-PCR.
2.2. Study Design
An open-label, non-randomized study was designed to examine the response of
BiosimCovex administered orally to healthy volunteers. This study aimed to examine
safety in terms of clinical effects and blood parameters, and to evaluate certain immune
responses at the baseline and at days 17, 34, and 60.
This study was conducted at a single center in Mumbai. The first volunteer was
enrolled on 10 June 2020, and the last volunteer’s visit was completed on 12 August 2020.
The study protocol, amendments, and informed consent forms (ICF) were reviewed and
approved by the Ethics Committee constituted by Homeopathy India Private Limited.
Written informed consent was obtained from each volunteer before the performance of any
study-specific procedures. This study was conducted in accordance with the guidelines
of the Good Clinical Practice and the ethical principles outlined in the Declaration of
Helsinki 2008.
2.3. Study Sample
This is a category ‘nosode’ preliminary HPT, with ten individuals proposed by the sci-
entific advisory board as appropriate for a Phase I trial. Ten healthy volunteers (
18–65 years
,
4 males and 6 females), with no known major untreated diseases, with normal routine
laboratory parameters during screening, were enrolled in the trial (Figure 1). The basic
demographic information is presented in Table 1. Subjects who currently had or were
recently diagnosed with COVID-19 infections were excluded from the study. Full inclusion
and exclusion criteria included the following:
Medicines 2023,10, 8 4 of 12
Medicines 2023, 10, x FOR PEER REVIEW 4 of 11
were recently diagnosed with COVID-19 infections were excluded from the study. Full
inclusion and exclusion criteria included the following:
Figure 1. Flow chart of an open-label, homeopathic pathogenetic trial for safety and the evaluation
of the immune response of BiosimCovex in healthy volunteers. ICF = Informed Consent Form.
Table 1. Demographics of Individual Subjects.
Subje
ct No.
Gender
(M.F)
Age
(Yrs)
Weight
(Kg)
WBC
(4.00–
10.00)
10
3
/µL)
RBC
(3.50–
5.50)
10
6
/uL
HGB
(11.0–
16.0)
g/dL
PLT
(100–
300)
10
3
/uL
Alkaline
Phospha
tase (53–
128) U/L
Bilirubi
n Direct
(0.00–
0.20)
mg/dL
Bilirubi
n Total
(0.00–
2.00)
mg/dL
Total
Protein
(6.40–
8.30)
g/dL
Albumi
n (3.5–
5.20)
g/dL
AST/GO
T (0.0–
35.0) U/L
ALT/GP
T (0.0–
45.0) U/L
C-
Reactive
Protein
(0.0–6.0)
mg/dL
Ferritin
(21.81–
274.66)
ng/mL
1 M 22 53 6.57 5.18 14 275 92 0.13 0.22 7.82 4.41 19.6 27.7 0.4 13.87
2 F 48 70.5 8.31 4.52 13.2 274 81 0.18 0.5 7.79 4.18 16.9 15.3 1.2 15.54
3 F 40 57.3 10.16 4.8 12 360 107 0.09 0.22 7.57 4.22 18.5 13.6 1.1 20.25
4 F 32 60 7.1 4.38 11.8 252 61 0.27 0.72 6.96 4.31 15.6 11.6 0.8 16.69
5 F 35 69 8.17 4.31 12.4 290 79 0.14 0.32 7.21 4.2 14.3 17.6 1.4 20.45
6 F 36 61 8.69 4.44 12 299 77 0.12 0.24 7.39 3.99 14.7 9.2 4.9 17.09
7 M 46 78 5.83 4.97 13.3 237 64 0.23 0.34 7.24 4.11 17.8 11.6 0.7 105.52
8 M 49 101.3 6.26 4.39 13.3 310 61 0.12 0.18 7.11 4.06 16.7 28.4 1.2 119.41
9 M 40 59.7 5.83 4.84 14.9 294 113 0.3 0.73 7.21 4.4 23.9 20.8 1.7 51.1
10 F 28 68 5.42 4.69 12.8 331 79 0.14 0.3 7.61 4.15 35.4 54.1 0.6 11.84
Mean 36.59 66.67 7.09 4.64 12.94 290.21 79.68 0.16 0.34 7.39 4.20 18.64 18.21 1.10 26.84
SD 8.80 13.89 1.54 0.29 0.98 36.15 18.05 0.07 0.20 0.29 0.14 6.29 13.38 1.29 40.32
Figure 1.
Flow chart of an open-label, homeopathic pathogenetic trial for safety and the evaluation of
the immune response of BiosimCovex in healthy volunteers. ICF = Informed Consent Form.
Medicines 2023,10, 8 5 of 12
Table 1. Demographics of Individual Subjects.
Subject
No.
Gender
(M.F)
Age
(Yrs)
Weight
(Kg)
WBC
(4.00–10.00)
103/µL)
RBC
(3.50–5.50)
106/uL
HGB
(11.0–16.0)
g/dL
PLT
(100–300)
103/uL
Alkaline
Phos-
phatase
(53–128)
U/L
Bilirubin
Direct
(0.00–0.20)
mg/dL
Bilirubin
Total
(0.00–2.00)
mg/dL
Total
Protein
(6.40–8.30)
g/dL
Albumin
(3.5–5.20)
g/dL
AST/GOT
(0.0–35.0)
U/L
ALT/GPT
(0.0–45.0)
U/L
C-Reactive
Protein
(0.0–6.0)
mg/dL
Ferritin
(21.81–274.66)
ng/mL
1 M 22 53 6.57 5.18 14 275 92 0.13 0.22 7.82 4.41 19.6 27.7 0.4 13.87
2 F 48 70.5 8.31 4.52 13.2 274 81 0.18 0.5 7.79 4.18 16.9 15.3 1.2 15.54
3 F 40 57.3 10.16 4.8 12 360 107 0.09 0.22 7.57 4.22 18.5 13.6 1.1 20.25
4 F 32 60 7.1 4.38 11.8 252 61 0.27 0.72 6.96 4.31 15.6 11.6 0.8 16.69
5 F 35 69 8.17 4.31 12.4 290 79 0.14 0.32 7.21 4.2 14.3 17.6 1.4 20.45
6 F 36 61 8.69 4.44 12 299 77 0.12 0.24 7.39 3.99 14.7 9.2 4.9 17.09
7 M 46 78 5.83 4.97 13.3 237 64 0.23 0.34 7.24 4.11 17.8 11.6 0.7 105.52
8 M 49 101.3 6.26 4.39 13.3 310 61 0.12 0.18 7.11 4.06 16.7 28.4 1.2 119.41
9 M 40 59.7 5.83 4.84 14.9 294 113 0.3 0.73 7.21 4.4 23.9 20.8 1.7 51.1
10 F 28 68 5.42 4.69 12.8 331 79 0.14 0.3 7.61 4.15 35.4 54.1 0.6 11.84
Mean 36.59 66.67 7.09 4.64 12.94 290.21 79.68 0.16 0.34 7.39 4.20 18.64 18.21 1.10 26.84
SD 8.80 13.89 1.54 0.29 0.98 36.15 18.05 0.07 0.20 0.29 0.14 6.29 13.38 1.29 40.32
Medicines 2023,10, 8 6 of 12
2.4. Inclusion Criteria
Age: 18–65 years, both males and females, healthy individuals with no major untreated
diseases and normal routine laboratory parameters during screening. Participants who
were informed of the nature of the study and were willing to give written informed consent
were enrolled in the study.
2.5. Exclusion Criteria
Any subject with a known disease or condition which might compromise the general
health or comorbid conditions, subjects who had been confirmed to have COVID-19 and
had been isolated for its treatment, subjects who had recently suffered and recovered
from COVID-19, persons with a known history of allergies and food hypersensitivity,
women during pregnancy, puerperium and breast-feeding, subjects who had participated
in another clinical trial during the last 6 months and with other conditions, which, in the
opinion of the investigators, could make the patient unsuitable for enrolment or could
interfere in their adherence to of the study protocol, were excluded from the study.
2.6. Study Method
Subjects were administered with six doses of nosode BiosimCovex 30C (pill size 30)
(Manufactured at Haffkine’s Institute-Batch no CoviNo-052020, Mfg. date: May 2020) as six
pills, twice daily, for three consecutive days. Pre and post examinations, such as physical
examinations, vital signs, and laboratory investigations were taken at baseline, on days 17,
34, and 60. The participants were monitored thoroughly. Based on the data in the source
documents and laboratory values, the safety results were reported. Samples were sent
to the testing laboratory within 30 min of collection at ambient conditions to avoid any
degradation. The central laboratory carefully processed the samples upon arrival.
2.7. Approvals
The trial protocol was reviewed by the Scientific Advisory Board and approved by an
Ethics Committee, named ‘Institutional Ethics Committee’, letter dated 27 May 2020. The
trial was registered at the Clinical Trial Registry of India (CTRI) with the trial registration
number: CTRI/2020/05/025496.
2.8. Study End Points and Statistical Assessments
The primary endpoint was the determination of the safety of the BiosimCovex, and the
secondary endpoint was the evaluation of the changes in the immune response parameters.
As the observations made after a specific time interval demonstrate the pre- and
post-treatment effects, and the data is continuous in nature, the t test for Paired Two Sample
for Means was explored. Statistical analyses were performed using Software SPSS (Version
1.0.0.1447 IBM Corp., Armonk, New York, United States). For a continuous dependent
variable IL-6, where the sample was collected at multiple visits, a significant difference
between observations in each visit was calculated by repeated measures ANOVA.
2.9. Safety Assessments
Safety assessments included the monitoring of subjects for any unexpected symptoms,
adverse events, serious adverse events, as assessed by predefined questionnaires, clinical
and laboratory investigation results, blood pressure, physical examination findings, and
general well-being, as assessed by questionnaire.
2.10. Safety Parameters
Complete Blood Count (CBC), C-Reactive Protein (CRP), Liver Function Test (LFT),
performed on the fully automated analyzer (XL-200), and Serum ferritin level, measured by
CMIA, and the chemiluminescent assay, were measured at baseline and day 17. The RT-PCR
tests for the laboratory verification of COVID-19 were not permitted by the government for
Medicines 2023,10, 8 7 of 12
healthy persons (due to the shortage of kits in the country during the study period). Instead,
the status of the COVID-19 IgG/IgM against the SARS-CoV-2 Spike Protein, measured
by enzyme-linked immunosorbent assay (ELISA) method (Index >1.1 positive, 0.9–1.1
borderline, and <0.9 negative), were investigated at baseline, days 17, 34, and 60, along
with the CBC and ferritin, to confirm that the subjects were not infected.
2.11. Immune Response Laboratory Method Parameters
An interferon–gamma (IFN-
γ
) assay was conducted by the Chemiluminescent assay
(CLIA) method, using a value less than 0.438 IU/ML as negative, and above that as positive;
this was measured at the baseline and day 17. In addition, Interleukin-6, using a value
greater than 7.00 pg/mL as positive and an electro chemiluminescent immunoassay (ECLIA,
Elecsys 2021 Modular Analytics E170 Cobas) method, was measured at the baseline and
days 17, 34 and 60. After initiating the study, and observing elevations of Interleukin-6, we
amended the protocol to also investigate the CD4 panel, measured at days 34 and 60.
3. Results
There were no serious or fatal adverse events during the study. The laboratory basic
biochemistry and liver function tests were not affected by BiosimCovex 30C (Table 2). The
P-value results of CBC, CRP, LFT, and the Serum ferritin level for all subjects at baseline,
compared to day 17, were unchanged, and in the normal reference range. The subjects
were closely monitored and did not show any abnormal clinical changes. Based on these
findings, no safety concerns were observed in the study population.
Table 2.
Basic laboratory parameters during the study course (t-Test, Paired two Sample for Means).
Sr No. Blood Parameters Baseline Day 17 pValue
1WBC (4.00–10.00) 103/µL) 7.234 7.529 0.2801
2RBC (3.50–5.50) 106/uL 4.652 4.208 0.2834
3 HGB (11.0–16.0) g/dL 12.97 12.97 1
4PLT (100–300)103/uL 292.2 276.4 0.0724
5
Alkaline Phosphatase (53–128) U/L
81.4 80.4 0.5897
6 Bilirubin Direct (0.00–0.20) mg/dL 0.172 0.231 0.0994
7 Bilirubin Total (0.00–2.00) mg/dL 0.377 0.573 0.0658
8 Total Protein (6.40–8.30) g/dL 7.391 7.28 0.1193
9 Albumin (3.5–5.20) g/dL 4.203 4.255 0.1920
10 AST/GOT (0.0–35.0) U/L 19.34 17.09 0.1679
11 ALT/GPT (0.0–45.0) U/L 20.99 16.74 0.2398
12
C-Reactive Protein (0.0–6.0) mg/dL
1.4 1.61 0.4992
13 Ferritin (21.81–274.66) ng/mL 39.176 38.803 0.9009
As this manuscript is focused on safety and potential immune response signals, symp-
toms that are usually cataloged during a homeopathic pathogenetic trial were cataloged
and are published elsewhere.
Regarding immune response, such as cytokine change, three subjects showed elevated
IL-6 on day 17 as compared to baseline, and all 10 subjects showed elevated IL-6 on
day 34. With the repetition of the IL-6 measure at day 60, the values of 9 subjects were
not different (returned) compared to the baseline. A significant difference between IL-6
observations, calculated by repeated measures ANOVA, was found highly significant
(
pvalue 0
). (Table 3) The subjects did not show any clinically untoward symptoms during
the time of the elevated IL-6 values. On the CD4 panel, the absolute CD4 count showed
Medicines 2023,10, 8 8 of 12
a significant elevation following the IL-6 timeframe on day 60, in comparison to day 34.
(Table 4). Individual supporting data presented in Supplementary Materials.
Table 3.
IL- 6 (0.00–7.00 pg/mL) changes after treatment in comparison with baseline, (RM-
ANOVA test).
Test (Unit) Visit Statistics n= 10 p-Value
IL- 6 (0.00–7.00 pg/mL) Visit 1 (Baseline) Mean (SD) 2.022 (0.794)
0.00094
Min, Max 1.5, 3.7
Visit 2 (Day 17) Mean (SD) 6.077 (2.845)
Min, Max 2.65, 12
Visit 3 (Day 34) Mean (SD) 29.032 (12.774) 0.00008
Min, Max 9.12, 44.6
Visit 4 (Day 60) Mean (SD) 5.829 (6.771) 0.11934
Min, Max 1.5, 24.26
Table 4. CD panel in individuals during the study (t-Test, Paired two samples for Means).
Test (Unit) Visit Statistics n= 10 p-Value
Absolute Lymphocyte count
(990.00–3150.00)/uL Visit 3 (Day 34) Mean (SD) 2198.7 (526.201)
0.25587
Min, Max 1563, 2854
Visit 4 (Day 60) Mean (SD) 2287.9 (502.911)
Min, Max 1699, 3004
Absolute CD4
(424.00–1509.00)/uL Visit 3 (Day 34) Mean (SD) 833.5 (280.553)
0.026799
Min, Max 505, 1453
Visit 4 (Day 60) Mean (SD) 900.8 (328.806)
Min, Max 526, 1520
Absolute CD8
(169.00–955.00)/uL Visit 3 (Day 34) Mean (SD) 652.3 (272.316)
0.2373
Min, Max 300, 1046
Visit 4 (Day 60) Mean (SD) 687.7 (293.047)
Min, Max 351, 1153
IFN-
γ
was measured at the baseline and day 17, [Baseline Mean (SD) 0.016 (0.025)
and day 17 Mean (SD) 0.13 (0.12)] and COVID-19 IgG and IgM measured at the baseline
(0.157/0.264), days 17 (0.352/0.207), 34 (0.220/0.231), or 60 (0.24/0.25), did not show any
activity indicating active exposure to SARS-CoV-2 virus during the dates measured.
4. Discussion
In conventional homeopathy, one conducts human pathogenetic trials (known as drug
provings), without carrying out any animal toxicity studies or safety studies, found in
precedents with earlier commonly-used nosodes such as Psorinum [
17
], Medorrhinum [
13
],
Tuberculinum [18], Carcinosin [19], Variolinum [13], and more.
The current trial of BiosimCovex in 30C potency studied in ten human subjects did not
elicit safety signals. It remains debatable within the homeopathic community whether the
conventional homeopathic pathogenetic trials should require acute, subacute, or chronic
toxicity in animal models, for examining the safety of nosodes; this is due to its poten-
Medicines 2023,10, 8 9 of 12
tized process which kills the original germs, if diluted, succussed and immersed in 90%
alcohol [20] during the process.
This human safety study was combined with the evaluation of the potential effects of
the nosode on the immune response. If the nosode was to be examined particularly for its
prophylactic potential during the pandemic, it is reasonable to expect an immune response
during the trial, and therefore this was evaluated. Being a pleiotropic cytokine, Interleukin-
6 (IL-6) is known to induce an antigen-specific immune response and inflammatory reaction,
activating the host defense mechanisms performing diverse functions. Studies of genetically
modified animal models suggest that IL-6 has a role in both the innate and adaptive immune
responses that protect the host from a variety of infections [21].
In this study, the mild elevation of IL-6 in all the subjects may be linked to an immune
response, as there were no corresponding clinical signs and symptoms of inflammatory
or pathological response. In addition, there was no increase in the C-Reactive Protein,
despite an increase in IL-6, suggestive of non-inflammatory and non-pathological responses,
thereby suggesting an immune response [22].
IL-6 levels, increasing by several tens or even hundreds of pg/mL, are observed
in chronic diseases, depending on severity. Values of more than 1000 pg/mL can occur
during septic shock or a cytokine storm and in severe cases [
8
]. The excessive or sustained
production of IL-6 is involved in various diseases [23].
The roles of IL-6 and IFN-
γ
are both important and well described, and both are
elevated greatly in COVID-19 patients [
24
]. Importantly, they are both investigated as
predictors of outcomes, including mortality. For example, IFN-
γ
is investigated as an
independent risk factor associated with mortality in the moderate and severe presentation
of COVID-19 [
25
]. Indeed, an emerging strategy is aimed directly at reversing the decline
in health status and lowering mortality rates in the acutely COVID-19 ill patient [26,27].
In our studies, unlike extreme changes, the IL-6 levels increased to an average of
five folds (statistically significant pvalue
≤
0.05). The current cytokine fluctuation was
observable on days 17 and 30, but not on day 60. IL-6 levels were reduced to baseline in
9 of the 10 subjects by day 60. Interestingly, the timing of elevated IL-6 found in these
subjects corresponds to the commonly observed timing of symptom production during a
conventional homeopathic pathogenetic trial [28].
In an influenza vaccination study, Mohanty et al. [
29
] had reported an increase in
cytokines, including IL-6, as an immune response following vaccination. An initial mild
rise in IL-6 may be considered a positive sign of immune response. They had also reported
a time-dependent elevation in intracellular cytokine production in monocytes with the
highest levels found at around day 28. In our study, subjects began showing an increase
in IL-6 on day 17, all having a significant rise by day 34, and subsequent decline by day
60. This corresponded to an increase in CD4 count by day 60, as compared to day 34 in
7 subjects
, thereby potentially suggesting the activation of innate immunity in response to
the nosode.
IL-6 is considered an important mediator of the immune response, particularly by
directly acting on CD4 T cells and determining their effector functions [
30
]. We observed
IL-6 elevation with a potential increase in the CD4 helper cell count at corresponding visits,
which is again an expressive marker of immunity.
Interestingly, this study observed a signal associated with the use of BiosimCovex,
potentially activating the IL-6 level in all the subjects and potentially influencing the innate
immune response by CD4 T cell differentiation in some. The T-cell-activation potential
of the nosode needs further studies. IFN-
γ
and immunoglobulins were not released at
any level.
While IL-6 stimulating the production of CRP [
31
] is well studied, we did not observe
any CRP [Baseline (1.4), day 17 (1.61), and day 34 (1.9)] elevation in this study. Lymphocyte
count was measured after IL-6 data demonstrated results; therefore, no baseline data is
available. With Lymphocyte counts remaining in the normal range, there could be no fear
that autoimmunity is triggered by this nosode.
Medicines 2023,10, 8 10 of 12
As discussed by Pérez-Galarza, et al., there are several key roles considered for CD4
in mediating the immune response when related to COVID-19, including originating
various phenotype T helper cells, such as Th1, to helping B cell create a long-lived antibody
response, to helping CD8 expand the primary immune response, to stimulating natural
killer cells’ cytotoxicity, to helping in viral clearance, and perhaps, memory CD4 helping to
prompt immune defense systems [32].
The clinical symptoms produced by the subjects were mild, short-lasting, and self-
limiting, and were carefully documented and reported in a separate homeopathic patho-
genetic trial article [
10
]. While a larger placebo-controlled HPT could be conducted in the
future to elicit more symptoms, the effects of the COVID-19 virus are well understood
due to their pandemic prevalence, and it may not be imperative to conduct symptomatic
HPT for its immediate prescribing as a prophylactic or therapeutic indication. Indeed,
in a related study, the authors conducted a randomized double-blind, placebo-controlled
feasibility study, evaluating the efficacy of homeopathic medicines in the prevention of
COVID-19 in a quarantined population. In 2233 quarantined individuals who were exposed
to people diagnosed with COVID-19, the subjects in the BiosimCovex arm reported signals
of efficacy, with a significantly smaller number of individuals testing COVID-19 positive
compared to the placebo group, as well as a shorter duration of illness compared to the
placebo [33].
IL-6 and CD4 changes were mild, but signal potential should be explored in a larger
follow-up trial. It is expected that these markers would show fewer changes than during
an infection or when elicited by a vaccine, though if these were seen in a large trial, it
could help the understanding of how homeopathic medicines might help in this public
health crisis. Interestingly, this finding matches previous results from other diseases. For
example, in a study on tularemia, Jonas presented results that were significantly better than
the placebo (22%), but not as effective as vaccinations (100%) [
34
]. If there is some level of
protection afforded by homeopathic prophylaxis, it may be useful for those who are unable
to tolerate vaccines or do not have access to them.
As epidemic and pandemic emergencies present with time constraints, immediate
prophylactic and therapeutic solutions may not wait for the lengthy process of new nosode
drug discovery. It is essential to develop faster, effective, yet safer methods of therapy; these
may include the development of nosodes from clinical samples or inactivated microbial
components, conducting safety studies, and evaluating specific immunological tests to
assess potential clinical usefulness. Should this strategy prove even partially effective,
it may fill the gap that occurs from the start of an epidemic until vaccine development,
production and distribution are underway. Further, with genomic variations, mutations,
and adoptions [
35
], it is imperative to search for alternate immune preventive or therapeu-
tic measures.
The limitations include Type 1 errors due to the small sample size in the trial and a lack
of a placebo-controlled arm. The lack of a placebo arm points us to the potential association
of the immune response, but not to causation. Another limitation is potentially the too
few immune markers being examined. Unfortunately, we added the CD4 count after we
observed a rise in IL-6, and, therefore, did not capture CD4 and Lymphocyte count baseline
levels. The study result is indicative of the immune response; however, a confirmatory
study should be repeated in the future in a larger sample size, with controls, and testing a
wider number of immune markers.
5. Conclusions
No safety concerns were observed in the small study population by consuming Biosim-
Covex. In this noncontrolled open-label study, while antibodies to COVID-19 remained
negative, a transient immune response may have been observed in elevated IL-6 in all
the subjects, with a corresponding increase in the CD4 count on day 60 in comparison to
day 34
. A larger placebo-controlled bridge study may be possible to explore the induction
of biochemical changes that may occur by ingestion of the nosode.
Medicines 2023,10, 8 11 of 12
Supplementary Materials:
The following supporting information can be downloaded at: https://
www.mdpi.com/article/10.3390/medicines10010008/s1, Table S1: IL-6 (0.00–7.00 pg/mL). Individual
subject changes after treatment in comparison with baseline, (RM-ANOVA test); Table S2: Individual
subject CD panel in individuals during the study (t-Test, Paired two Sample for Means).
Author Contributions:
P.H., manuscript writing and statistical analysis; G.T., research coordination,
manuscript writing; S.V., assistance in nosode development, protocol, and manuscript review; R.S.,
principal investigator, nosode development, manuscript writing. All authors have read and agreed to
the published version of the manuscript.
Funding: No external funding was received for this study.
Institutional Review Board Statement:
The study protocol, amendments, and informed consent
forms (ICF) were reviewed and approved by the Ethics Committee constituted by Homeopathy In-dia
Private Limited. This study was conducted in accordance with the guidelines of the Good Clinical
Practice and the ethical principles outlined in the Declaration of Helsinki 2008.
Informed Consent Statement:
Written informed consent was obtained from each volunteer before
the performance of any study-specific procedures.
Data Availability Statement: Not applicable.
Acknowledgments:
Our sincere thanks to the members of the Scientific Advisory Board of Life
Force Foundation Trust, members of the Institutional Ethics Committee, AYUSH mentor Shubhadha
Chiplunkar (Former director, ACTREC, TMC) and Rakesh Rawal (Gujarat University, Ahmedabad,
India) for valuable inputs in the data evaluation. Our thanks go to Mala Vazirani and the team at
Transasia Biomedicals for providing laboratory facilities. Inputs on statistical evaluation by Shwetal
Talele (Purdue University, West Lafayette, IN, USA) and Anton Igonin (Göttingen, Germany) are
acknowledged. We extend our thanks to Sandeepan Mukherjee from Haffkine institute, Mumbai, for
providing facility and support in the development of the nosode.
Conflicts of Interest:
Paul Herscu has no conflict of interest. Gitanjali Talele works with Biosilia but
had no role in data analysis or interpretation. Rajesh Shah was granted a patent for the nosode used
in this study.
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