<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0206</schemaVersion>

    <documentType>3</documentType>

    <periodOfReport>2020-04-22</periodOfReport>

    <noSecuritiesOwned>0</noSecuritiesOwned>

    <issuer>
        <issuerCik>0001724344</issuerCik>
        <issuerName>Entasis Therapeutics Holdings Inc.</issuerName>
        <issuerTradingSymbol>ETTX</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001080014</rptOwnerCik>
            <rptOwnerName>Innoviva, Inc.</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>1350 OLD BAYSHORE HIGHWAY SUITE 400</rptOwnerStreet1>
            <rptOwnerStreet2></rptOwnerStreet2>
            <rptOwnerCity>BURLINGAME</rptOwnerCity>
            <rptOwnerState>CA</rptOwnerState>
            <rptOwnerZipCode>94010</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>0</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>1</isTenPercentOwner>
            <isOther>0</isOther>
            <officerTitle></officerTitle>
            <otherText></otherText>
        </reportingOwnerRelationship>
    </reportingOwner>

    <nonDerivativeTable>
        <nonDerivativeHolding>
            <securityTitle>
                <value>Common Stock</value>
            </securityTitle>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>1322510</value>
                    <footnoteId id="F1"/>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </nonDerivativeHolding>
    </nonDerivativeTable>

    <derivativeTable>
        <derivativeHolding>
            <securityTitle>
                <value>Warrants to Purchase Common Stock</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>2.50</value>
            </conversionOrExercisePrice>
            <exerciseDate>
                <value>2020-04-22</value>
            </exerciseDate>
            <expirationDate>
                <value>2025-04-22</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>1322510</value>
                    <footnoteId id="F1"/>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeHolding>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">As reflected in the Schedule 13D filed by Innoviva, Inc. (the &quot;Reporting Person&quot;) with the U.S. Securities and Exchange Commission on April 23, 2020 (the &quot;Schedule 13D&quot;), in connection with an initial closing that occurred on April 22, 2020 pursuant to a securities purchase agreement, dated as of April 12, 2020 (the &quot;Purchase Agreement&quot; and, such closing, the &quot;Initial Closing&quot;), by and between Entasis Therapeutics Holdings Inc. (the &quot;Issuer&quot;) and the Reporting Person, the Reporting Person acquired 1,322,510 shares of common stock of the Issuer (&quot;Shares&quot;) and warrants to purchase an additional 1,322,510 Shares (&quot;Warrants&quot;) as set forth in this Form 3. As a result of the Initial Closing, the Reporting Person beneficially owns 2,645,020 Shares.  The Schedule 13D filed by the Reporting Person is incorporated by reference in this Form 3; any description herein of the Schedule 13D is qualified in its entirety by reference to the Schedule 13D so filed by the Reporting Person.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Geoffrey Hulme</signatureName>
        <signatureDate>2020-04-23</signatureDate>
    </ownerSignature>
</ownershipDocument>
